Source:http://linkedlifedata.com/resource/pubmed/id/20932077
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| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
2
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| pubmed:dateCreated |
2010-10-11
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| pubmed:abstractText |
The paucity of appropriate reagents for serologic typing of the Diego blood group antigens has prompted the development of a real-time PCR and melting curve analysis for Diego blood group genotyping. In this study, we phenotyped 4326 donor blood samples for Di(a) using semiautomated equipment. All 157 Di(a+) samples were then genotyped by PCR using sequence-specific primers (PCR-SSP) for DI*02 because of anti-Di(b) scarcity. Of the 4326 samples, we simultaneously tested 160 samples for Di(a) and Di(b) serology, and DI*01 and DI*02 by PCR-SSP and by real-time PCR. We used the same primers for Diego genotyping by real-time PCR and PCR-SSP. Melting curve profiles obtained using the dissociation software of the real-time PCR apparatus enabled the discrimination of Diego alleles. Of the total samples tested, 4169 blood donors, 96.4 percent (95% confidence interval [CI], 95.8-96.9%), were homozygous for DI*02 and 157, 3.6 percent (95% CI, 3.1%-4.2%), were heterozygous DI*01/02. No blood donor was found to be homozygous for DI*01 in this study. The calculated DI*01 and DI*02 allele frequencies were 0.0181 (95% CI, 0.0173-0.0189) and 0.9819 (95% CI, 0.9791-0.9847), respectively, showing a good fit for the Hardy-Weinberg equilibrium. There was full concordance among Diego phenotype results by PCR-SSP and real-time PCR. DI*01 and DI*02 allele determination with SYBR Green I and thermal cycler technology are useful methods for Diego determination. The real-time PCR with SYBR Green I melting temperature protocol can be used as a rapid screening tool for DI*01 and DI*02 blood group genotyping.
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| pubmed:language |
eng
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| pubmed:journal | |
| pubmed:citationSubset |
IM
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| pubmed:chemical | |
| pubmed:status |
MEDLINE
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| pubmed:issn |
0894-203X
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| pubmed:author | |
| pubmed:issnType |
Print
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| pubmed:volume |
26
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| pubmed:owner |
NLM
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| pubmed:authorsComplete |
Y
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| pubmed:pagination |
66-70
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| pubmed:meshHeading |
pubmed-meshheading:20932077-Alleles,
pubmed-meshheading:20932077-Anion Exchange Protein 1, Erythrocyte,
pubmed-meshheading:20932077-Blood Donors,
pubmed-meshheading:20932077-Blood Group Antigens,
pubmed-meshheading:20932077-Blood Grouping and Crossmatching,
pubmed-meshheading:20932077-Female,
pubmed-meshheading:20932077-Heterozygote,
pubmed-meshheading:20932077-Humans,
pubmed-meshheading:20932077-Male,
pubmed-meshheading:20932077-Reverse Transcriptase Polymerase Chain Reaction,
pubmed-meshheading:20932077-Sensitivity and Specificity
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| pubmed:year |
2010
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| pubmed:articleTitle |
Application of real-time PCR and melting curve analysis in rapid Diego blood group genotyping.
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| pubmed:affiliation |
Immunohematology Division, Fundacao Pro-Sangue Hemocentro de Sao Paulo, Assistant Professor Hematology Department, University of Sao Paulo Medical School, Av. Dr. Eneas Carvalho de Aguiar, 155-1 andar, Sao Paulo, SP-Brazil CEP 05403-000, Brazil.
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| pubmed:publicationType |
Journal Article
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