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Article | 14-October-2020

Enzyme and DTT treatment of adherent RBCs for antibody identification by a solid phase immunoassay system

Treatment of RBCs with protease enzymes or dithiothreitol (DTT) causes denaturation of several RBC antigens and is regularly used in antibody identification. In this study, we have standardized enzyme and DTT treatment of adherent RBCs in the magnetic-mixed passive hemagglutination assay (M-MPHA) for antibody identification. We have also tried drying these treated RBCs. The optimal enzyme and DTT treatment conditions for intact adherent RBCs were determined, in addition to the optimal condition

Toyohiro Tamai, Toshio Mazda

Immunohematology, Volume 18 , ISSUE 4, 114–119

Article | 14-October-2020

Loss of enzyme-sensitive antigens due to the presence of leukocytes, neomycin sulfate, and LISS

Previous studies have shown that RBCs with residual WBCs stored in LISS and neomycin sulfate develop characteristics associated with enzyme-treated RBCs. During a mass screening program to antigen type donor RBCs, we observed that the Fya antigens on a RBC sample from an in-house panel became non-detectable with anti-Fya after incubation overnight in Diluent 2 from Micro Typing Systems, Inc. (MTS, Pompano Beach, FL). In response to this observation, we initiated an investigation to determine

Randall W. Velliquette, Paula Howard, Harry Malyska, Marion E. Reid

Immunohematology, Volume 19 , ISSUE 4, 109–111

Short Communication | 26-August-2016

Basidiospore and Protoplast Regeneration from Raised Fruiting Bodies of Pathogenic Ganoderma boninense

Ganoderma boninense, a phytopathogenic white rot fungus had sought minimal genetic characterizations despite huge biotechnological poten­tials. Thus, efficient collection of fruiting body, basidiospore and protoplast of G. boninense is described. Matured basidiocarp raised under the glasshouse conditions yielded a total of 8.3 × 104 basidiospores/ml using the low speed centrifugation technique. Mycelium aged 3-day-old treated under an incubation period of 3 h in lysing enzyme from

Nisha Thopla Govender, Maziah Mahmood, Idris Abu Seman, Wong Mui-Yun

Polish Journal of Microbiology, Volume 65 , ISSUE 3, 383–388

Article | 18-October-2020

Comparison of human platelet antigen (HPA)-1a typing by solid phase red cell adherence to HPA-1 allotypes determined by allelespecific restriction enzyme analysis

Phenotype results for human platelet antigen (HPA)-1 by CaptureP®‚ (Immucor, Inc., Norcross, GA) solid phase red cell adherence (SPRCA) were compared to results of allele-specific restriction enzyme analysis (ASRA) for the determination of HPA-1 allotype. Because the expression of HPA-1a and HPA-1b is determined by a single nucleotide substitution of thymine‘cytosine at position 196 of the gene encoding membrane glycoprotein (GP)-IIIa, it is possible to distinguish the alternate

Michael J. McGann, Jo L. Procter, Junichi Honda, Kazuhiko Matsuo, David F. Stroncek

Immunohematology, Volume 16 , ISSUE 2, 68–73

Review | 01-December-2019

Warm autoadsorption with enzyme-treated red blood cells

Patients demonstrating warm autoantibody specificity present serologic challenges for laboratory staff performing antibody identification in the blood bank. Autoantibody can be removed from plasma or serum by adsorption onto autologous red blood cells (RBCs) provided the patient has not been transfused in the previous 3 months. The adsorption process can be enhanced by enzyme pretreatment of autologous RBCs.

Farai Tsimba-Chitsva, Susanne Bishop, Kelly Kezeor

Immunohematology, Volume 28 , ISSUE 3, 88–90

Article | 16-February-2021

Use of trypsin in serologic investigation

reliable commercial source of trypsin, additional proteolytic enzymes such as papain and ficin were later identified as useful. Enzyme treatment is thought to decrease the negative charge between RBCs by the removal of sialic acid. It was soon recognized that the treatment of RBCs with chemicals (e.g., dithiothreitol [DTT] and 2-aminoethylisothiouronium bromide) or enzymes (e.g., ficin, papain, trypsin, and α-chymotrypsin) alters the expression of RBC antigens differently. The change in antibody

A. Novotny

Immunohematology, Volume 35 , ISSUE 4, 145–148

Article | 14-October-2020

Evaluation of a new solid-phase immunoassay for alloantibody detection using bromelin-treated and untreated red blood cells

The enzyme test is used to detect certain antibodies or facilitate antibody identification. This study compares antibody reactivity with bromelin-treated red blood cells (RBCs) and untreated RBCs using a newly developed solid-phase immunoassay. The reactivity of irregular antibodies was tested by a magnetic-mixed passive hemagglutination assay (M-MPHA). In addition, antibody reactivity was tested with dried stroma of bromelin-treated RBCs and untreated RBCs (M-MPHA-Dry). Rh antibodies were

Toyohiro Tamai, Toshio Mazda

Immunohematology, Volume 17 , ISSUE 1, 17–21

Article | 17-November-2020

Application of the proteolytic enzyme papain in routine platelet serology

The use of proteolytic enzymes is well established in red cell serology. These enzymes modify some antigen structures and remove sialic acid from the red cell membrane. Enzyme-sensitive structures have also been identified on the platelet membrane. The effect of papain, a proteolytic enzyme used widely in red cell serology, on the detection of various platelet alloantibodies was examined to determine its usefulness in platelet serology. Antisera with the specificities anti-HPA-la, -2b, -3a, -4a

John A.G. Lown, Brian J. Dale

Immunohematology, Volume 11 , ISSUE 4, 140–142

Article | 14-October-2020

Quantitation of red cell–bound IgG, IgA, and IgM in patients with autoimmune hemolytic anemia and blood donors by enzymelinked immunosorbent assay

This paper describes an enzyme immunoassay for the quantitative determination of IgG, IgA, and IgM immunoglobulins on RBCs. Ether eluates made from RBCs were followed by an enzyme-linked immunosorbent assay of immunoglobulin concentration. Calibration curves were derived from immunoglobulin standards and the number of molecules of each isotype per RBC was calculated. The assay was carried out in 200 healthy blood donors and 62 patients with warm autoimmune hemolytic anemia (AIHA), two of them

Antonio A. Bencomo, Martha Díaz, Yalile Alfonso, Odalys Valdés, María E. Alfonso

Immunohematology, Volume 19 , ISSUE 2, 47–53

Article | 09-November-2020

Measurement of red blood cell-bound C3b and C3d using an enzyme-linked direct antiglobulin test

Complement has a complex role in immune mediated red blood cell (RBC) destruction and usually induces extravascular hemolysis of C3bcoated RBCs by erythrophagocytosis and by acting synergistically with cell-bound immunoglobulins. A sensitive two-stage enzyme-linked direct antiglobulin test (ELDAT) was developed and used to measure RBC-bound C3b and C3d in 120 healthy adult individuals and in 60 patients suffering from a variety of conditions, including warm- and cold-type autoimmune hemolytic

J.D. Bellamy, D.J. Booker, N.T. James, R. Stamps, R.J. Sokol

Immunohematology, Volume 13 , ISSUE 4, 123–131

Article | 10-April-2021

An automated approach to determine antibody endpoint titers for COVID-19 by an enzyme-linked immunosorbent assay

directed against the RBD correlate with neutralizing antibody levels, demonstrating that anti-RBD levels may provide a useful surrogate for SARS-CoV-2 antibody activity.14 This finding is especially important when considering that, although neutralizing assays are often considered the gold standard for assessing antiviral antibody activity, these approaches can be difficult to establish as high-throughput tests, making it difficult to implement this approach clinically.15 Using an enzyme-linked

A.D. Ho, H. Verkerke, J.W. Allen, B.J. Saeedi, D. Boyer, J. Owens, S. Shin, M. Horwath, K. Patel, A. Paul, S.-C. Wu, S. Chonat, P. Zerra, C. Lough, J.D. Roback, A. Neish, C.D. Josephson, C.M. Arthur, S.R. Stowell

Immunohematology, Volume 37 , ISSUE 1, 33–43

research-article | 30-November-2019

Ultrastructure of Hirschmanniella diversa early-stage infection in browning rhizomes of Indian lotus

Shigeru Uematsu, Tetsuo Yabu, Mitsuyoshi Yao, Takayuki Kurihara, Hironori Koga

Journal of Nematology, Volume 52 , 1–9

Research paper | 06-February-2018

Impact of recurrent hypoglycemic stress on hindbrain A2 nerve cell energy metabolism and catecholamine biosynthesis: modulation by estradiol

It is unclear if habituation of hindbrain A2 metabolo-sensory neurons to recurrent insulin-induced hypoglycemia (RIIH) correlates with estradiol-dependent adjustments in energy metabolism that favor positive energy balance. Laser-microdissected A2 cells from estradiol- or oil-implanted ovariectomized female rats were analyzed by Western blot to assess effects of three prior daily insulin injections on basal and hypoglycemic patterns of catecholamine biosynthetic enzyme dopamine-beta-hydroxylase

Pratistha Tamrakar, Karen P. Briski

Acta Neurobiologiae Experimentalis, Volume 77 , ISSUE 1, 31–44

Article | 14-October-2020

One thousand seventy antibodies detected only by a 2-stage papain test: wanted and unwanted positive reactions

Carmen Martin-Vega, Dolores Castella, Joan Cid, Marta Panadés

Immunohematology, Volume 17 , ISSUE 4, 122–124

original-paper | 03-September-2019

Purification, Characterization and Inhibition of Alanine Racemase from a Pathogenic Strain of Streptococcus iniae

developed resistance against many potential antibiotics (Tavares et al. 2018). As such, additional efforts for developing more effective vaccines and antibiotics are necessary steps for circumventing the threat of its infection (Saavedra et al. 2004). Alanine racemase (Alr; E.C. is an enzyme that catalyzes the interconversion of L-alanine and D-alanine using a pyridoxal 5-phosphate (PLP) as a cofactor (Tassoni et al. 2017). It provides D-alanine for the synthesis of peptidoglycan of the


Polish Journal of Microbiology, Volume 68 , ISSUE 3, 331–341

Review | 27-December-2020

A review: the enzyme-linked antiglobulin test (ELAT) and its applications for reference laboratories

Ruth Mougey

Immunohematology, Volume 5 , ISSUE 3, 69–78

research-article | 08-July-2020

The inhibitory effects of bile acids on catalytic and non-catalytic functions of acetylcholinesterase as a therapeutic target in Alzheimer’s disease

, AChE inhibition by various drugs, such as donepezil, rivastigmine and galantamine, are used to improve symptoms of AD pathophysiology (Anand and Singh, 2013). AChE which belongs to the α/β hydrolase fold family is an ACh-hydrolyzing enzyme in the synapse and neuromuscular junction that terminates cholinergic impulses in nerves (Holmquist, 2000). Despite the catalytic activity, AChE participates in multiple biological functions, such as neuronal cell differentiation, dendrite and axon formation

Leila Sadeghi, Reza Yekta, Gholamreza Dehghan

Acta Neurobiologiae Experimentalis, Volume 80 , ISSUE 2, 108–116

Article | 14-October-2020

Low-incidence MNS antigens associated with single amino acid changes and their susceptibility to enzyme treatment

Marion E. Reid, Jill Storry

Immunohematology, Volume 17 , ISSUE 3, 76–81

Report | 16-March-2020

The polymorphism nt 76 in exon 2 of SC is more frequent in Whites than in Blacks

The Scianna blood group system comprises seven antigens encoded by alternative forms of SC. The SC gene also has two polymorphisms in the leader sequence, at nucleotides 54 (C/T, silent) and 76 (C/T, 26His/Tyr) in exon 2, which are not involved in expression of blood group antigens. The nucleotide change at position 76 has an NlaIII restriction enzyme site; thus, DNA samples from 100 Caucasians and 100 African Americans were analyzed for the SC nucleotide 76 change. DNA from Caucasian and

Akiko Fuchisawa, Christine Lomas-Francis, Kim Hue-Roye, Marion E. Reid

Immunohematology, Volume 25 , ISSUE 1, 18–19

original-paper | 04-June-2020

Performance Evaluation of Different Commercial Serological Kits for Diagnosis of Acute Hepatitis E Viral Infection

hepatitis E cases mainly depends on the serological detection of anti-HEV antibodies (Dreier and Juhl 2014). However, equivalence, sensitivity, and specificity in the results of the HEV Enzyme-linked Immunosorbent Assay (ELISA) kits tend to differ between manufacturers, leading to discrepancies in the rates of anti-HEV antibodies among different populations (Herremans et al. 2007; Drobeniuc et al. 2010), together with the HEV genome heterogeneity, and the different antigenic structure of HEV proteins


Polish Journal of Microbiology, Volume 69 , ISSUE 2, 217–222

Article | 15-February-2021

ZZAP treatment of red blood cells

Principle ZZAP is a mixture of a sulfhydryl reagent (dithiothreitol [DTT]) and a proteolytic enzyme (papain or ficin). It was first described by Branch and Petz in their 1982 article in the American Journal of Clinical Pathology.1 ZZAP is used to dissociate IgG and complement from red blood cells (RBCs), an action that neither reagent can achieve alone. According to Branch and Petz, it is believed that ZZAP “reduces interchain disulfide linkages, increasing exposure of the IgG polypeptides to

S.I. Marckwardt

Immunohematology, Volume 35 , ISSUE 1, 9–10

Article | 09-November-2020

A modified PCR-RFLP genotyping method demonstrates the presence of the HPA-4b platelet alloantigen in a North American Indian population

the HPA-4 antigen system does not involve a common naturally occurring restriction enzyme site. This paper describes a new genotyping method for HPA-4 (polymerase chain reaction–restriction fragment length polymorphism [PCR-RFLP]) that involves restriction enzyme digestion of PCR-amplified genomic DNA using a modified PCR primer to create an artificial TaqI restriction site that is present in the HPA-4a but not in the HPA-4b DNA sequence. The HPA-4 PCR-RFLP method was validated by testing a

Alexander P. Reiner, Gayle Teramura

Immunohematology, Volume 13 , ISSUE 2, 37–43

Research Article | 15-February-2020

Polypyrrole Based Nano-Biosensor for Sensing Low Concentrations of Hydrogen Peroxide

This paper discuss about a novel technique of developing a nano-structured polypyrrole biosensor for measurement of very low concentrations of hydrogen peroxide in liquid media. The proposed fabrication method is very effective in growing a nano-structured conducting polymer layer on a planar conducting substrate. In addition enzyme loading was done under a high electric field of 1000V/m. The developed sensor provides a liner range of 0-200µM of hydrogen peroxide and a measurement

D.M.G. Preethichandra, M. Onoda

International Journal on Smart Sensing and Intelligent Systems, Volume 7 , ISSUE 5, 1–4

Article | 14-December-2020

Paternity evaluation in the presence of splits and crossreactive antigens

Evaluation of paternity (alleged father, mother, and child) can range from a strightforward resolution to a complex problem that cannot be resolved without family studies. We present a case of disputed paternity in which tests for crossreactive groups (CREGs) and antigen subtypes (splits) within the human leukocyte antigen (HLA) system could not be used confidently to prove or disprove paternity. Further analysis, red cell enzyme tests, enabled a final verdict and confirmed the current

Robert W. Gutendorf, Kamala Balakrishnan, David L. Taylor, Kelly Cox

Immunohematology, Volume 7 , ISSUE 2, 43–45

original-paper | 28-January-2020

Banana Peels: A Promising Substrate for the Coproduction of Pectinase and Xylanase from Aspergillus fumigatus MS16

biological research (Otero and Nielsen 2010). Previously, enzymatic digestion of several LC materials has been reported such as corncob (Kahar et al. 2010), Kraft paper-mill sludges (Kang et al. 2010), and sugarcane bagasse (Buaban et al. 2010). It suggests that LC materials can be saccharified using an enzyme(s) and can provide a cheaper source of raw materials for many industries. However, the selection of an LC substrate for industrial exploitation mainly depends on local agricultural practices, as


Polish Journal of Microbiology, Volume 69 , ISSUE 1, 19–26

Short Communication | 27-September-2017

Clonal Analysis of Clinical and Environmental Pseudomonas aeruginosa Isolates from Meknes Region, Morocco

From 123 clinical and environmental Pseudomonas aeruginosa isolates, 24 strains were selected for their similar antibioresistance, virulence and biofilm formation profiles, to examine their diversity and occurrence of clones within two hospitals and different natural sites in Meknes (Morocco). Pulsed-field gel electrophoresis, using DraI enzyme, didn’t reveal a close relationship between clinical and environmentalisolates nor between strains of the two hospitals. 19 genotypes were

Itto Maroui, Abouddihaj Barguigua, Asmae Aboulkacem, Hanane Elhafa, Khadija Ouarrak, Mohammed Sbiti, Lhoussain Louzi, Mohammed Timinouni, Abdelhaq Belhaj

Polish Journal of Microbiology, Volume 66 , ISSUE 3, 397–400

Article | 06-December-2020

Effect of enzymes on and chemical modifications of high-frequency red cell antigens

Enzyme or chemical modification of intact red cells results in the destruction of some blood group antigens. The pattern of reactions of an antibody with red cells treated with various proteinases, with sialidase, and with the disulfide bond-reducing agent 2-aminoethylisothiouronium bromide (AET) can aid in antibody identification. This information can prove particularly beneficial with antibodies to antigens of very high frequency, where antigen-negative cells may be difficult to obtain

Geoff Daniels

Immunohematology, Volume 8 , ISSUE 3, 53–57

Report | 01-December-2019

Detection and identification of plateletassociated alloantibodies by a solidphase modified antigen capture enzymelinked immunosorbent assay method and its correlation to platelet refractoriness in multiplatelet concentrate–transfused patients

are transfused with multiple units of leukodepleted platelet concentrates, such as those with hemato-oncologic diseases and bone marrow failure syndromes. The method used was solidphase modified antigen capture enzyme-linked immunosorbent assay. Platelet refractoriness was assessed by measuring the corrected count increment at 1 and 24 hours after transfusion.

Neelesh Jain, Ravi Shankar Sarkar, Joseph Philip

Immunohematology, Volume 30 , ISSUE 3, 123–125

Article | 17-November-2020

Identification of the Tcb allele of the Cromer blood group gene by PCR and RFLP analysis

antigen-negative cells and typing sera. In blacks, about 5 percent of individuals carry the rare Tcb Cromer allele. We have shown that the presence ofthe low-incidence Tcb allele can be detected by polymerase chain reaction (PCR) amplification of a fragment of the gene encoding DAF, followed by allele-specific restriction enzyme digestion.

Manisha Udani, Nicole Anderson, Neeraja Rao, Marilyn J. Telen

Immunohematology, Volume 11 , ISSUE 1, 1–4

Article | 15-February-2021

Donath-Landsteiner test

intravascular hemolysis of RBCs when the autoantibody dissociates. This test shows a positive result when hemolysis is visibly seen in the test system (Fig. 1). Fig. 1 Indirect Donath-Landsteiner test. Reagents/Supplies Reagents Supplies Indirect DL test Fresh pooled normal sera 50% suspension of washed P+ RBCs Glass test tubes 0°C ice bath 37°C heat block or incubator Centrifuge Enzyme-treated indirect DL test 1% papain-treated P+ RBCs Two-stage indirect DL test Group O RBCs

M. Kilty, T.S. Ipe

Immunohematology, Volume 35 , ISSUE 1, 3–6

Article | 16-October-2019

Separation of multiple antibodies by adsorption with allogeneic red blood cells

. Routinely, antibody identification practices in a blood bank comprise the testing of a patient’s plasma against reagent RBCs using standard agglutination and indirect antiglobulin methods.1 There are, however, instances when identification of multiple antibodies may be complicated and require additional serologic methods. When the presence of multiple antibodies is suspected, several methods—including neutralization of patient’s plasma, titration, elution, chemical or enzyme treatment of reagent RBCs

E.M. Ekema

Immunohematology, Volume 33 , ISSUE 4, 155–158

Case report | 29-December-2020

Evaluation of a complement-dependent anti-Jka by various sensitization and detection methodologies: a case report

A 79-year-old woman with a diagnosis of lower gastrointestinal bleeding was found to have a complement-dependent anti-Jka in her serum. The anti-Jka was evaluated by the antiglobulin technique with polyspecific, anti-C3, and anti-IgG antihuman globulin (AHG). A variety of sensitization and detection methods were used, including the prewarmed saline technique, enzyme treatment of test cells, a low-ionic additive solution (LISS), 22 percent albumin, Polybrene, and an increased serum/cell ratio

E. Nicole DeLong

Immunohematology, Volume 4 , ISSUE 3, 59–63

Article | 29-December-2020

Immunoglobulin A (IgA) levels in blood products and plasma derivatives

lgA was measured by radial immunodiffusion and enzyme-linked immunosorbent assay techniques. Blood products which consistently contained IgA less than 0.05 mg/dL (the present definition for IgA deficiency used by the American Red Cross Rare Donor Registry) were from IgA-deficient donors, deglycerolized red blood cells (RBCs) prepared with an extra wash cycle, and RBCs washed using a total volume of approximately 1300 mL of 0.9 percent sodium chloride.

Susan M. Fox, Linda M. Stavely-Haiber

Immunohematology, Volume 4 , ISSUE 1, 5–9

Article | 18-October-2020

Quantitation of red cell-bound immunoglobulins and complement in lymphoma patients

M. Podberezin, A. Levina, L. Romanova, O. Margolin, O. Nasibov, A.V. Pivnik

Immunohematology, Volume 16 , ISSUE 4, 147–153

Article | 18-October-2020

From kill to overkill: 100 years of (perhaps too much) progress

Peter D. Issitt

Immunohematology, Volume 16 , ISSUE 1, 18–25

Article | 18-May-2020

The gene encoding the I blood group antigen: review of an I for an eye

Unlike most blood group antigen pairs, the I and i antigens are not antithetical (produced by allelic pairs) but, rather, they are reciprocal. The I antigen is formed by the action of an enzyme (a glycosyltransferase), which adds branches onto the i antigen. Thus, branched I antigen is formed at the expense of its precursor, the linear i antigen. The antigens are present on all blood cells and have a wide tissue distribution. Soluble I antigen is found in milk, saliva, and amniotic fluid, and a

Marion E. Reid

Immunohematology, Volume 20 , ISSUE 4, 249–252

Article | 30-November-2020

Autoimmune hemolysis following transfusion: a mimicking autoanti-D in a D- patient with alloanti-D

An 80-year-old group O, D- (rr) female with anti-C, -D, -E, and -Fya received four units of crossmatch-compatible red blood cells (RBCs). The direct antiglobulin test (DAT) was negative. Two weeks later, jaundice, dark urine, a 16% drop in hematocrit (Hct), a 20% reticulocyte count, and absent haptoglobin occurred. During the next month, her DAT was positive with anti-IgG and -C3d. Acid eluates, which repeatedly showed anti-D specificity, were nonreactive with enzyme-treated D- RBCs. Adsorption

Walter H. Dzik, Joyce Blank, Paula Lutz, Thomas G. Hirose, Christine Lomas-Francis, Marilyn Moulds

Immunohematology, Volume 10 , ISSUE 4, 117–119

Original Paper | 30-June-2018

Screening and Identification of Trichoderma Strains Isolated from Natural Habitats with Potential to Cellulose and Xylan Degrading Enzymes Production

A total of 123 Trichoderma strains were isolated from different habitats and tested for their ability to degrade cellulose and xylan by simple plate screening method. Among strains, more than 34 and 45% respectively, exhibited higher cellulolytic and xylanolytic activity, compared to the reference strain T. reesei QM 9414. For strains efficiently degrading cellulose, a highest enzyme activity was confirmed using filter paper test, and it resulted in a range from 1.01 to 7.15 FPU/ml. Based on


Polish Journal of Microbiology, Volume 67 , ISSUE 2, –

research-article | 30-November-2020

Hydrolytic Enzymes Producing Bacterial Endophytes of Some Poaceae Plants

in many areas of the industry because they are more stable, cheaper, and can be obtained in large amounts by fermentation methods (Singh et al. 2016). Examples of industrial areas affected by discoveries of these enzymes include detergent agents, leather processing, degradation of xenobiotic compounds, food processing (bakery, meat, dairy, fruit, and vegetable products), pharmaceuticals (synthesis of pharmaceutical intermediates), biofuels (low–energy ethanol production process), and other enzyme


Polish Journal of Microbiology, Volume 70 , 1–8

Original Paper | 07-June-2016

Enzymes Involved in Naproxen Degradation by Planococcus sp. S5

, 3,4-dihydroxybenzoic acid or vanillic acid as growth substrates, the degradation of 21.5%, 71.71%, 14.75% and 8.16% of naproxen was observed respectively. It was shown that the activity of monooxygenase, hydroxyquinol 1,2-dioxygenase, protocatechuate 3,4-dioxygenase and protocatechuate 4,5-dioxyegnase in strain S5 was induced after growth of the strain with naproxen and 4-hydroxybenzoate. Moreover, in the presence of naproxen activity of gentisate 1,2-dioxygenase, enzyme engaged in 4

Danuta Wojcieszyńska, Dorota Domaradzka, Katarzyna Hupert-Kocurek, Urszula Guzik

Polish Journal of Microbiology, Volume 65 , ISSUE 2, 177–182

Article | 09-November-2020

Semiautomation of platelet HPA-1a phenotyping by SPRCA and ELISA

An enzyme-linked immunosorbent assay (ELISA) and solid phase red cell adherence assay (SPRCA) were assessed for platelet HPA-1a typing in U well microplates. Both methods were partially automated by the use of the Tecan RSP 8051ID robotic sampler and the SLT 400 ATC plate reader with Soft2000 software. Pretreatment of the adherent platelets with chloroquine diphosphate or citric acid enabled anti-HPA-1a, even when contaminated with HLA class 1 antibodies, to be used for typing. Of 675 antenatal

Lionel A. Mohabir, Lynne Porter

Immunohematology, Volume 13 , ISSUE 2, 44–48

Original Paper | 04-September-2018

RNA Quality Control Using External Standard RNA

mRNA, inhibition to DNA polymerase, and degradation of mRNA for determining the RNA quality using standard RNA. It would be possible to know yield of mRNA and inhibition of the enzyme reaction by adding standard RNA before RNA extraction and looking at standard RNA loss. Degradation was evaluated by comparing the differences in the 3’ and 5’ regions of the RNA. In our study, it was demonstrated that in the crude extract of Saccharomyces cerevisiae, degradation was comparatively higher at the 3’ end


Polish Journal of Microbiology, Volume 67 , ISSUE 3, 347–353

Original Paper | 10-December-2018

A Special Risk Group for Hepatitis E Infection: The First Record of North Cyprus

husbandry, veterinary work or butchery). Enzyme-linked immunoassays were used to detect anti-HEV IgG and IgM in the blood samples. The prevalence of anti-HEV IgG antibodies was 3.0% (12/400), while the prevalence of anti-HEV IgM antibodies was 0.25% (1/400). The prevalence of anti-HEV IgG amongst the samples received from females was approximately 2.5-fold higher than samples received from males (2.4%). Anti-HEV IgG was detected amongst 7% of animal husbandry workers and amongst 2% of veterinarians and


Polish Journal of Microbiology, Volume 67 , ISSUE 4, 525–528

research-article | 30-November-2018

Active and inactive forms of biotin synthase occur in Heterodera glycines

biochemistry, development, and defenses (Hussey, 1989; Smant et al., 1998; Lambert et al., 1999; Davis et al., 2000). Not all nematode secretory proteins, however, originate from the esophageal glands and some of the genes that encode secretory proteins can act as either virulence or avirulence genes. An esophageal gland-secreted protein/enzyme chorismate mutase (CM) is thought to suppress host defense by lowering the levels of chorismate-derived compounds required for plant defense (Lambert et al., 1999

Khee Man Kwon, Sadia Bekal, Leslie L. Domier, Kris N. Lambert

Journal of Nematology, Volume 51 , 1–12

original-paper | 04-June-2020

Lactobacillus fermentum JX306 Restrain D-galactose-induced Oxidative Stress of Mice through its Antioxidant Activity

stool consistency, immune modulation, and antagonism towards the pathogens. Another attractive feature of the LAB is their antioxidant capacity. Increasing experimental evidence indicates that probiotic LAB exerts beneficial antioxidative effects by ROS scavenging, chelating transition metal ions, and activating certain enzyme activities. Therefore, using LAB to scavenge the excess of free radicals, inhibit oxidative damage, and prevent the related restrictive diseases can be a potential treatment


Polish Journal of Microbiology, Volume 69 , ISSUE 2, 205–215

Article | 26-October-2020

DNA from urine sediment or buccal cells can be used for blood group molecular genotyping

with cotton wool swabs. DNA, prepared using a commercial kit, was subjected to polymerase chain reaction amplification and followed by digestion with the appropriate restriction enzyme. Genotyping was performed for three alleles encoded by polymorphic genes on three different chromosomes, namely KEL1/KEL2, JKA/JKB, and FYA/FYB. Genotyping results were compared to the results of typing performed on red blood cells using standard hemagglutination techniques. Results given by samples freshly collected

Marion E. Reid, Maria J. Rios, Kevin L. Cash, Annie M. Strupp, Joan M. Uehlinger

Immunohematology, Volume 15 , ISSUE 2, 61–65

original-paper | 01-May-2021

Screening of Human Immunodeficiency Virus (HIV) among Newly Diagnosed Tuberculosis Patients in Eastern Sudan

collected from each patient, then serum samples were separated (Tognon et al. 2020) and investigated for HIV antibodies by using fourth-generation enzyme-linked immunosorbent assay (ELISA) according to the manufacturer’s instructions. Data was analyzed by IBM SPSS Statistics for Windows, Version 20 (Armonk, NY: IBM Corp) and iNZight (The University of Auckland New Zealand). Chi-square test was used to test the p-value, and it was deemed significant if it was less than 0.05. Ethical approval for this


Polish Journal of Microbiology, Volume 70 , ISSUE 2, 201–206

Original Research | 18-July-2017

Diversity of Root-knot Nematodes Associated with Tubers of Yam (Dioscorea spp.) Established Using Isozyme Analysis and Mitochondrial DNA-based Identification

identified using enzyme phenotyping (esterase and malate dehydrogenase) and mitochondrial DNA (mtDNA) NADH dehydrogenase subunit 5 (Nad5) barcoding. Examination of 48 populations revealed that yam tubers were infested by Meloidogyne incognita (69%), followed by M. javanica (13%), M. enterolobii (2%), and M. arenaria (2%). Most of the tubers sampled (86%) were infected by a single species, and multiple species of RKN were detected in 14% of the samples. Results of both identification methods revealed the

Yao A. Kolombia, Gerrit Karssen, Nicole Viaene, P. Lava Kumar, Nancy de Sutter, Lisa Joos, Danny L. Coyne, Wim Bert

Journal of Nematology, Volume 49 , ISSUE 2, 177–188

Research paper | 06-February-2018

Assessment of type 1 and type 3 deiodinase expression levels in depressive disorders

. We aimed to investigate the levels of DIO1 and DIO3 in the patients suffering from recurrent depressive disorders (rDD). Data collected from 91 rDD patients and 105 healthy controls were analyzed. The diagnoses were made based on the ICD-10 criteria (F33.0–F33.8). The expression levels of DIO1 and DIO3 were estimated using the polymerase chain reaction method and the enzyme-linked immunosorbent assay (ELISA). The expression of DIO1 on mRNA/protein levels in the rDD patients was reduced in

Elżbieta Gałecka, Anna Kumor-Kisielewska, Agata Orzechowska, Michael Maes, Paweł Górski, Janusz Szemraj

Acta Neurobiologiae Experimentalis, Volume 77 , ISSUE 3, 225–235

Original Paper | 15-March-2016

Characterization and Optimization of Biosynthesis of Bioactive Secondary Metabolites Produced by Streptomyces sp. 8812

®ZYM test were analyzed and genetic analysis made. Phylogenetic analysis of Streptomyces sp. 8812 revealed that its closest relative is Streptomyces capoamus JCM 4734 (98%), whereas sequence analysis for 16S rRNA gene using NCBI BLAST algorithm showed 100% homology between these two strains. Biosynthetic processes, mycelium growth and enzyme inhibitory activities of these two strains were also compared.

Aleksandra Rajnisz, Adam Guśpiel, Magdalena Postek, Joanna Ziemska, Anna Laskowska, Daniel Rabczenko, Jolanta Solecka

Polish Journal of Microbiology, Volume 65 , ISSUE 1, 51–61

original-paper | 30-November-2018

New Insight into Genotypic and Phenotypic Relatedness of Staphylococcus aureus Strains from Human Infections or Animal Reservoirs

superfamily. These toxins are responsible for staphylococcal food poisoning, but they also affect some immune system cells, with further consequences (Zhang et al. 2017). S. aureus strains also secrete a variety of enzymes. Proteases include a vast group of secreted enzymes, such as aureolysin, serine proteases, and staphopains, which are engaged in the evasion of complement-mediated bacterial killing (Miedzobrodzki et al. 2002; Sabat et al. 2008; Otto 2014). Nuclease, another extracellular enzyme


Polish Journal of Microbiology, Volume 68 , ISSUE 1, 93–104

original-paper | 27-March-2019

Mycosynthesis of Size-Controlled Silver Nanoparticles through Optimization of Process Variables by Response Surface Methodology

formation of AgNPs and reduction of silver nitrate to its colloidal state (Jogee et al. 2017) as illustrated (Fig. 3A). High enzyme activity was observed throughout the experiments, with the highest being 179.15 nmol/h/ml (Fig. 3B). All the OS experimental runs produced monodispersed AgNPs. At higher substrate and metal salt concentration, the enzyme activity seemed delayed and larger sized AgNPs were formed; which could be due to unavailability of functional groups that were responsible for carrying


Polish Journal of Microbiology, Volume 68 , ISSUE 1, 35–42

Article | 17-November-2020

Loss of the Knops blood group system antigens from stored blood

Complement receptor type one (CR1) is a polymorphic glycoprotein, present on red blood cells (RBCs), that carries the Knops blood group system antigens. Since Knops system antigens can vary in strength, we investigated whether CR1 deteriorated upon storage, thus affecting Knops blood group system antigen reactivity. Units of whole blood were collected in CPDA-1 and evaluated at day 0 and day 35 for antigen strength, using routine serologic techniques. CR1 was quantitated by an enzyme-linked

Joann M. Moulds, L. Lee Brown, Elizabeth Brukheimer

Immunohematology, Volume 11 , ISSUE 2, 46–50

Article | 06-December-2020

Cytomegalovirus antibody screening on the Olympus PK7100

(LAT). There were four discrepant results in the 1,495 donor plasmas tested by the two methods. Two of the three samples that were positive by the LAT and negative by the OHA test were also negative with the IMx enzyme-linked immunosorbent assay (ELISA) and complement fixation test (CFT). The OHA failed to detect antibodies to CMV in a sample that was positive by the IMx ELISA, CFT, and LAT. The IMx ELISA, which was regarded as the arbiter of true positivity, yielded a sensitivity and specificity

LioneI A. Mohabir

Immunohematology, Volume 8 , ISSUE 2, 41–43

Article | 01-April-2020

A single base insertion of the 4-α-galactosyltransferase gene led to the deficiency of Gb3 biosynthesis

of Gb3/CD77 antigen on the cell surface was evaluated by flow cytometry and by immunochemical techniques. All individuals with the p phenotype were found to have a single base insertion (A4GALT/insC) at the same nucleotide position. Neither the transfectant cells with a mutant gene (A4GALT/insC) of donor origin or those with a synthesized mutant gene (A4GALT/insC-Mu) expressed Gb3 antigen indicating that the presence of A4GALT/insC diminished the A4GALT enzyme activity. In addition, an allele

Mitsunobu Tanaka, Naoko Yamashita, Junko Takahashi, Fumiya Hirayama, Yoshihiko Tani, Hirotoshi Shibata

Immunohematology, Volume 22 , ISSUE 1, 23–29

research-article | 30-November-2019

Effect of Heterodera schachtii female age on susceptibility to three fungal hyperparasites in the genus Hyalorbilia

from the edge of the fungal block, and plates were incubated in the dark at 23°C. Healthy surface-sterilized females were placed on water agar plates as controls. After 3 days, 25 parasitized or control females from each plate were transferred to the center of water agar plates, 5 females per plate, and 5 plates per fungal strain or control. Plates were incubated in the dark at 23°C for 6 days, after which females were removed from plates and used for enzyme assays. Hatched J2 were rinsed from

J. Smith Becker, J. Borneman, J. O. Becker

Journal of Nematology, Volume 52 , 1–12

research-article | 26-March-2021

Aminoguanidine ameliorates ovariectomy-induced neuronal deficits in rats by inhibiting AGE-mediated Aβ production

Dan Di Zhang, Yan Gang Wang, Chun Yan Liu, Ze Hou Wang, Yue Fen Wang

Acta Neurobiologiae Experimentalis, Volume 81 , ISSUE 1, 10–20

Review | 25-March-2020

The O2 allele: questioning the phenotypic definition of an ABO allele

There are three main alleles in the ABO blood group system, A, B, and O.  The former two alleles encode glycosyltransferases resulting in the wild-type A and B phenotypes, whereas the latter allele does not encode a functional enzyme owing to a frameshift polymorphism in the majority of cases.  Thus the group O phenotype is the absence of A or B sugars.  More than 15 years ago the O2 allele was described; this allele did not feature the usual crippling 261delG polymorphism, which

Mark H. Yazer, Martin L. Olsson

Immunohematology, Volume 24 , ISSUE 4, 138–147

Article | 03-November-2020

Use of the MAIEA assay to demonstrate that Fy3 is on the same glycoprotein as Fy6, Fya, and Fyb

The monoclonal-antibody immobilization of erythrocyte antigens (MAIEA) assay is a technique that detects trimolecular complexes formed by a human antibody and a mouse monoclonal antibody with specific red cell epitopes. This enzyme-linked immunoadsorbent assay test gives a positive reaction when two different epitopes on the same membrane protein are separately recognized by human and mouse antibodies. In this study, the MAIEA test was used to determine if the Duffy system antigen Fy3 is on the

Jaw-Lin Tzeng, Roger Dodd, Delores Mallory

Immunohematology, Volume 14 , ISSUE 3, 113–116

Article | 27-August-2018


more efficient in samples with mycelium immobilized on a polypropylene foam, what probably was associated with increased enzyme activity of the strains, as well as enhancement of the contact of the dye with the mycelium. Strain L3 respectively removed 100% (mycelium immobilized) of the dye after 24h and 95.8% (mycelium suspended) of the dye after 96h. For complete removal of the dye the immobilized biomass of strain L3 needs 24 hours of incubation, and L1 48h. Strain L1 completely removed the color


Architecture, Civil Engineering, Environment, Volume 10 , ISSUE 1, 137–145

Review Paper | 06-April-2016

Anticonvulsant therapy in brain-tumor related epilepsy

seizures does not differ substantially from that applied to epilepsies from other etiologies. Therefore, the choice of an AED is based, above all, on tolerability and pharmacokinetic interactions with chemotherapeutic drugs. Levetiracetam is recommended by many authors as first-line therapy in brain tumor-related epilepsy. Due to the possibility of interactions, the combination of enzyme-inducing AEDs and chemotherapeutic drugs, is usually not recommended as a first choice. Currently there is no

Walter Fröscher, Timo Kirschstein, Johannes Rösche

Journal of Epileptology, Volume 24 , ISSUE 1, 41–56

Article | 01-April-2020

An alloantibody to a highprevalence MNS antigen in a person with a GP.JL/Mk phenotype

. Immunoblotting showed the presence of monomer and dimer forms of a GP(A-B) hybrid and an absence of GPA and GPB. Sequencing of DNA and PCR-RFLP using the restriction enzyme RsaI confirmed the presence of a hybrid GYP(AB). The patient’s antibody was determined to be anti-EnaFR. She is the first person reported with the GP.JL phenotype associated with a deletion of GYPA and GYPB in trans to GYP.JL.

John Ratliff, Susan Veneman, Joan Ward, Christine Lomas-Francis, Kim Hue-Roye, Randall W. Velliquette, Laima Sausais, Twilla Maldonado, Janet Miyamoto, Yolanda Martin, David Slater, Marion E. Reid

Immunohematology, Volume 23 , ISSUE 4, 146–149

review-article | 26-March-2021

Putative mechanism of neurological damage in COVID-19 infection

longer incubation period. Therefore, differential controls should be adopted as soon as possible. The incubation period for the virus is around 6.4 days (ranges from 0 to 24 days) (Wu et al., 2020). Role of the ACE2 receptor in COVID-19 infection We used three different databases for analyses, protein expression and localization. The human angiotensin-converting enzyme 2 (ACE2) is a zinc metallopeptidase containing 805 amino acids, the gene has been mapped to the X chromosome (Xp22) and it has 82

Cindy Bandala, José Luis Cortes-Altamirano, Samuel Reyes-Long, Eleazar Lara-Padilla, Ian Ilizaliturri-Flores, Alfonso Alfaro-Rodríguez

Acta Neurobiologiae Experimentalis, Volume 81 , ISSUE 1, 69–79

Report | 06-November-2019

Drug-induced immune neutropenia/ agranulocytosis

, sulfamethoxazoletrimethoprim, β-lactam antibiotics, clozapine, levamisole, and vancomycin. Assays used for detection of neutrophil drug-dependent antibodies (DDAbs) include flow cytometry, monoclonal antibody immobilization of granulocyte antigens, enzyme-linked immunosorbent assay, immunoblotting, granulocyte agglutination, and granulocytotoxicity. However, testing for neutrophil DDAbs is rarely performed owing to its complexity and lack of availability. Mechanisms proposed for DIIN have not been rigorously studied

Brian R. Curtis

Immunohematology, Volume 30 , ISSUE 2, 95–101

original-paper | 30-November-2018

The Influence of Temperature and Nitrogen Source on Cellulolytic Potential of Microbiota Isolated from Natural Environment

. hormones or vitamins), improvement of nitrogen and phosphorus nutrition of plants, and biological protection against pathogenic organisms. The aim of this study was to isolate from the environmental samples the microorganisms with high cellulolytic activity, which are able to grow and synthesize of the enzyme in a wide spectrum of temperature from cheap nitrogen source and may be used as a component of biopreparations. Experimental Materials and Methods Strains isolation. The strains were obtained


Polish Journal of Microbiology, Volume 68 , ISSUE 1, 105–114

Research Article | 22-May-2019


roots of host plants release strigolactones, which stimulate germination and branching of spores of arbuscular fungi. As a result, the fungi synthesize molecular signals, i.e. chitooligosaccharides (COs) and lipochitooligosaccharides (LCOS), called MycF factors. Thanks to the development of molecular biology techniques the probable cascade of events during the recognition of fungal MycF factor by the host-plant has been outlined. The enzyme 3-hydroxy-3-methylglutaryl coenzyme A reductase 1 (HMGR1

Katarzyna Jas, Urszula Małolepsza

Postępy Mikrobiologii - Advancements of Microbiology, Volume 56 , ISSUE 3, 275–281

Article | 03-November-2020

Autoimmune hemolytic anemia caused by warm-reacting IgM-class antibodies

confirmed as IgM by their ability to rebind to normal red blood cells (RBCs) after elution; the absence of small increases in RBC-bound IgG and IgA was shown by a sensitive enzyme-linked antiglobulin test. Patient 1 was a 64-year-old female with non-Hodgkin’s lymphoma, with a hemoglobin of 50 g/L and haptoglobin of < 0.1 g/L. Direct antiglobulin tests were positive for IgM, C3d, and C3c; only IgM was present in an eluate. The serum contained a weak autoantibody at 37°C and tests for

R.J. Sokol, D.J. Booker, R. Stamps, S. Sobolewski, A.P. Haynes

Immunohematology, Volume 14 , ISSUE 2, 53–58

Original Paper | 27-September-2017

Metallo-Beta-Lactamase Producing Pseudomonas aeruginosa in a Healthcare Setting in Alexandria, Egypt

isolates were identified using standard microbiological methods and were tested for their antimicrobial susceptibility patterns using single disc diffusion method according to the Clinical and Laboratory Standards Institute recommendations. Thirty P. aeruginosa isolates were randomly selected and tested for their MBL production by both phenotypic and genotypic methods. Diagnostic Epsilometer test was done to detect metallo-beta-lactamase enzyme producers and polymerase chain reaction test was done to

Amani F. Abaza, Soraya A. El Shazly, Heba S.A. Selim, Gehan S.A. Aly

Polish Journal of Microbiology, Volume 66 , ISSUE 3, 297–308

research-article | 25-June-2021


wirusa i błoną komórki. Głównymi receptorami komórkowymi wykorzystywanymi przez koronawirusy człowieka są: aminopeptydaza N (APN – aminopeptidase N) rozpoznawana przez HCoV-229E, konwertaza angiotensyny typ 2 (ACE2 – angiotensin-converting enzyme 2) używana przez wirusy SARS-CoV, SARS-CoV-2 i HCoV-NL63, oraz peptydaza dwupeptydylowa IV (DPP4 – dipeptidyl-peptidase IV) swoista dla MERS-CoV [20, 39, 61]. Nawet niewielka zmiana w obrębie RBD białka S może zmieniać jego powinowactwo do receptora. Yang i

Jolanta Bratosiewicz-Wąsik, Tomasz J. Wąsik

Advancements of Microbiology – Postepy Mikrobiologii, Volume 60 , ISSUE 2, 121–135

original-paper | 22-October-2019

Salmonella-Infected Aortic Aneurysm: Investigating Pathogenesis Using Salmonella Serotypes

protein extraction. Cytokines determination. Quantitative determination of IL-1β (R&D Systems, DLB50), IL-12p40 (Blue-Gene Biotech, Shanghai, China, E01I0045), IL-12p35 (BlueGene Biotech, Shanghai, China, E01I0030), and interferon (IFN)-α (PBL Interferon Source, 41100) was performed through enzyme-linked immunosorbent assay (ELISA) in culture supernatants according to the manufacturer’s protocol. The experiments were performed in triplicate and presented as mean ± SD. Protein extraction and Western


Polish Journal of Microbiology, Volume 68 , ISSUE 4, 439–447

research-article | 16-April-2020

De novo transcriptome sequencing and analysis of Anisakis pegreffii (Nematoda: Anisakidae) third-stage and fourth stage larvae

extracted from each APL3 and APL4 samples as mentioned above; 1 μl of total RNA extracted using Trizol (Sigma. USA) was mixed with 2 μl of 5 X gDNA Eraser Buffer, 1 μl of gDNA Eraser, 6 μl of RNase-free dH2O, and incubated at 42˚C for 2 min. Then, cDNA was synthesized by adding 4 μl of 5 X PrimeScript™ Buffer 2, 1 μl of PrimeScript™ RT Enzyme Mix I, 1 μl of RT Primer Mix, 4 μl of RNase-free dH2O, and incubated at 37˚C for 15 min. The selected A. pegreffii gene sequences and the internal control primers

U-Hwa Nam, Jong-Oh Kim, Jeong-Ho Kim

Journal of Nematology, Volume 52 , 1–16

Report | 01-December-2019

SC*994C>T causes the Scnull phenotype in Pacific Islanders and successful transfusion of Sc3+ blood to a patient with anti-Sc3  

*994C>T change introduces a restriction enzyme cleavage site for Tsp45I, and polymerase chain reaction (PCR) products from exon 12 were subjected to this PCR–restriction fragment length polymorphism (RFLP) assay. The five samples had the variant SC*994T/T. One sample, from a first cousin of one Marshallese proband, was heterozygous for SC*1514C/T (in the 3′ untranslated region); the other four samples were SC*1514C/C (consensus sequence). Samples from white donors (n = 100) and

Marion E. Reid, Kim Hue-Roye, Randall W. Velliquette, Kathleen Larimore, Sue Moscarelli, Nicolas Ohswaldt, Christine Lomas-Francis

Immunohematology, Volume 29 , ISSUE 2, 69–72

Case report | 09-October-2019

Hematologic complications in a patient with Glycine soja polyagglutination following fresh frozen plasma transfusion

Polyagglutination is a rare and underdiagnosed condition, characterized by agglutination of red blood cells (RBCs) with almost all ABO-compatible adult sera. Polyagglutination can occur when a cryptantigen is exposed on RBCs via microbial enzyme activity. Because nearly all adults naturally produce antibodies against cryptantigens, transfusion of plasma can cause unexpected hemolysis and hematologic complications, such as thrombocytopenia and disseminated intravascular coagulation, in patients

Ryan P. Jajosky, Lloyd O. Cook, Elizabeth Manaloor, James F. Shikle, Roni J. Bollag

Immunohematology, Volume 33 , ISSUE 2, 51–55

Report | 06-November-2019

Drug-induced immune thrombocytopenia: incidence, clinical features, laboratory testing, and pathogenic mechanisms

candidate drug resulted in recurrent thrombocytopenia. Flow cytometry testing for DDAbs can be useful in confirmation of a clinical diagnosis, and monoclonal antibody enzyme-linked immunosorbent assay testing can be used to determine the platelet glycoprotein target(s), usually GPIIb/IIIa or GPIb/IX/V, but testing is not widely available. Several pathogenic mechanisms for DIIT have been proposed, including hapten, autoantibody, neoepitope, drug-specific, and quinine-type drug mechanisms. A recent

Brian R. Curtis

Immunohematology, Volume 30 , ISSUE 2, 55–65

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