Dataset Information

HNA-1, -3, -4, and -5 genotyping using multiplex PCR among southern Thais: Developing continual HNA-1 null detection.

ABSTRACT: BACKGROUND:Antibodies against human neutrophil antigens (HNAs) are involved in various clinical conditions including transfusion-related acute lung injury and auto/alloimmune neutropenia. We aimed to determine HNA-1, -3, -4, and -5 frequencies among southern Thais using multiplex PCR and to develop HNA-1 null detection. METHODS:Samples obtained from 427 southern Thai blood donors were genotyped HNA-1, -3, -4, and -5 using multiplex PCR and compared their allele frequencies with those previously reported in Thai populations. HNA-1 null samples were tested by newly developed PCR-SSP and PCR-RFLP and confirmed by DNA sequencing. RESULTS:The frequencies of FCGR3B*01, FCGR3B*02, FCGR3B*03, SLC44A2*01, SLC44A2*02, ITGAL*01, and ITGAL*02 among southern Thais differed from other Thai populations, except ITGAM*01 and ITGAM*02 frequencies. Two samples without specific fragments of FCGR3B*01, FCGR3B*02, and FCGR3B*03 tested by multiplex PCR were confirmed by PCR-RFLP to identify FCGR3B deficiency (HNA-1 null). Moreover, to reduce test steps, the newly developed PCR-SSP for FCGR3B deficiency was validated and tested in all samples and the results were in agreement with DNA sequencing. CONCLUSIONS:This was the first study to report HNA-1, -3, -4, and -5 frequencies among southern Thais. The indeterminate results of multiplex PCR for HNA-1 genotyping led to establish HNA-1 null detection using PCR-SSP, which is simple, convenient and cost-effective and can be used to identify FCGR3B deficiency.

SUBMITTER: Intharanut K

PROVIDER: S-EPMC6430332 | biostudies-literature | 2019 Jan

REPOSITORIES: biostudies-literature

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HNA-1, -3, -4, and -5 genotyping using multiplex PCR among southern Thais: Developing continual HNA-1 null detection.

Intharanut Kamphon K Sasikarn Wiradee W Mitundee Supattra S Nathalang Oytip O

Journal of clinical laboratory analysis 20180813 1

<h4>Background</h4>Antibodies against human neutrophil antigens (HNAs) are involved in various clinical conditions including transfusion-related acute lung injury and auto/alloimmune neutropenia. We aimed to determine HNA-1, -3, -4, and -5 frequencies among southern Thais using multiplex PCR and to develop HNA-1 null detection.<h4>Methods</h4>Samples obtained from 427 southern Thai blood donors were genotyped HNA-1, -3, -4, and -5 using multiplex PCR and compared their allele frequencies with th ...[more]

PMID: 30105756

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Project description:BackgroundAntibodies to human neutrophil alloantigens (HNA) are involved in the pathophysiology of several clinical conditions including transfusion-related acute lung injury (TRALI), alloimmune and autoimmune neutropenia, and febrile nonhemolytic transfusion reactions leading to neutropenia. The cognate antigens are polymorphic structures expressed on several glycoproteins on the neutrophils, i.e., antigens HNA-1a, -1b, -1c, and -1d on Fc-γ-receptor IIIb; HNA-2 on CD177; HNA-3a and -3b on choline transporter-like protein 2; HNA-4a and -4b on CD11b/αM subunit of the αMβ2-integrin (CD11b/CD18, Mac-1, CR3); and HNA-5a and -5b on αL-subunit (CD11a) of the αLβ2 integrin (CD11a/CD18), leukocyte function associated molecule (LFA)-1. Currently, there is a lacuna of diagnostic methods for detection of HNA in India. This study aimed to determine the HNA frequencies in Indians, estimate the risk of alloimmunization, and prepare typed neutrophil panels, which can be used to detect HNA antibodies in neutropenia cases.Material and methodsEDTA blood samples were collected from random 1,054 blood donors. HNA-2 was phenotyped on fresh EDTA samples using FITC labelled monoclonal anti-CD177 by flowcytometry. HNA-1 (FCGR3B) genotyping was carried out by DNA sequencing and PCR-RFLP. Antigens of HNA-3 (SLC44A2) and HNA-5 (ITGAL) were genotyped by PCR-RFLP using TaqαI and Bsp1286I restriction enzymes, respectively, while HNA-4 (ITGAM) was genotyped by PCR-SSP.ResultsAllele frequencies of FCGR3B*01, FCGR3B*02, and FCGR3B*03 were found to be 0.433, 0.444, and 0.087, respectively. FCGR3B*01+*02+*03- was the most common genotype (33.78%). Ten individuals showed deficiency of FCGR3B individuals, while 23 showed hyperexpression, i.e., FCGR3B*01+*02+*03+. FCGR3B*04and *05 occurred with a frequency of 0.002 and 0.024. HNA-2 was found to be a high frequency antigen occurring in 98.8% population. Four percent individuals showed atypical expression of CD177 on their neutrophils. Allele frequencies of SLC44A2*01 and SLC44A2*02were 0.812 and 0.188, respectively, and that of ITGAM*01, ITGAM*02, ITGAL*01, and ITGAL*02 were 0.9546, 0.0454, 0.2372, and 0.7628, respectively.ConclusionThis is the first study in India to report the frequencies of HNA among blood donors. Typed neutrophil panels identified in the present study will enable us to investigate suspected cases of immune neutropenia in future.

Dataset Information

HNA-1, -3, -4, and -5 genotyping using multiplex PCR among southern Thais: Developing continual HNA-1 null detection.

Publications

HNA-1, -3, -4, and -5 genotyping using multiplex PCR among southern Thais: Developing continual HNA-1 null detection.

Similar Datasets

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