PMID- 24697294
OWN - NLM
STAT- MEDLINE
DCOM- 20141121
LR  - 20181202
IS  - 1423-0410 (Electronic)
IS  - 0042-9007 (Linking)
VI  - 107
IP  - 2
DP  - 2014 Aug
TI  - Establishment of reference panel for human platelet antigen genotyping.
PG  - 166-70
LID - 10.1111/vox.12149 [doi]
AB  - BACKGROUND AND OBJECTIVES: Human platelet antigens (HPAs) are platelet-specific 
      alloantigens associated with polymorphisms of platelet surface glycoproteins 
      (GPs), and they can induce alloantibodies when individuals lacking a particular 
      polymorphism are exposed to them via pregnancy or transfusion. Immune responses 
      to HPAs are involved in the pathogenesis of several clinical syndromes. HPA 
      genotyping is therefore important for clinical diagnosis and laboratory research. 
      This study aims to establish a reference panel for HPA genotyping. MATERIALS AND 
      METHODS: Genomic DNA extracted from human blood was used as the template for 
      amplifying HPA (1a-5a and 15a) gene fragments using specific primers. The 
      amplified products were cloned into pGM-T vectors, which were transformed into 
      competent TOP10 cells. After clone screening and amplification, the plasmids were 
      extracted and sequenced. Next, the gene fragments HPA-1b-5b and 15b were obtained 
      by site-directed mutagenesis using the corresponding HPA-1a-5a and 15a plasmids 
      as template DNA. RESULTS: We successfully constructed reference plasmids for HPA 
      genotyping with HPA-1a-5a, 15a, HPA-1b-5b and 15b. The DNA sequences were 
      consistent with those published in GenBank. CONCLUSION: Obtaining reference DNA 
      for low-frequency HPAs is very difficult, and the successful construction of 
      reference plasmids for the six HPA systems may solve this problem. Establishment 
      of this panel has laid the foundation for future research on HPA genotyping.
CI  - (c) 2014 International Society of Blood Transfusion.
FAU - Li, R S
AU  - Li RS
AD  - Shanghai Institute of Blood Transfusion, Shanghai Blood Center, Shanghai, China.
FAU - Qiao, Z L
AU  - Qiao ZL
FAU - Ling, B
AU  - Ling B
FAU - Lu, P
AU  - Lu P
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20140402
PL  - England
TA  - Vox Sang
JT  - Vox sanguinis
JID - 0413606
RN  - 0 (Antigens, Human Platelet)
SB  - IM
MH  - Antigens, Human Platelet/*genetics
MH  - Base Sequence
MH  - Blood Platelets/immunology
MH  - Genotype
MH  - Genotyping Techniques/*standards
MH  - Humans
MH  - Mutagenesis, Site-Directed
MH  - Plasmids/genetics
MH  - Platelet Transfusion
MH  - Polymorphism, Restriction Fragment Length
MH  - Polymorphism, Single-Stranded Conformational
MH  - Reference Standards
MH  - Sequence Analysis, DNA
OTO - NOTNLM
OT  - alloantibody
OT  - human platelet antigen
OT  - reference panel
OT  - site-directed mutagenesis
OT  - thrombocytopenia
EDAT- 2014/04/05 06:00
MHDA- 2014/12/15 06:00
CRDT- 2014/04/05 06:00
PHST- 2013/10/17 00:00 [received]
PHST- 2014/02/28 00:00 [revised]
PHST- 2014/03/04 00:00 [accepted]
PHST- 2014/04/05 06:00 [entrez]
PHST- 2014/04/05 06:00 [pubmed]
PHST- 2014/12/15 06:00 [medline]
AID - 10.1111/vox.12149 [doi]
PST - ppublish
SO  - Vox Sang. 2014 Aug;107(2):166-70. doi: 10.1111/vox.12149. Epub 2014 Apr 2.