An analysis of the reason for HLA-C allele dropout in five samples by sequence-based typing / 中华医学遗传学杂志
Chinese Journal of Medical Genetics
;
(6): 562-566, 2009.
Article
in Chinese
| WPRIM
| ID: wpr-287374
ABSTRACT
<p><b>OBJECTIVE</b>To analyze the possible reason for HLA-C allele dropout in routine sequence-based typing (SBT) and improve the accuracy of HLA-C SBT test.</p><p><b>METHODS</b>A total of 620 randomly selected samples from healthy voluntary blood donors in Shenzhen were typed at HLA-C locus by sequence-based typing using the AlleleSEQR HLA-C plus sequence-based typing kit. Samples with no full match result were subjected to cloning and haplotype sequencing of the full-length HLA-C gene. If no novel mutations were found, samples were then retyped, using our self-designed PCR primer pair and PCR conditions replacing the AlleleSEQR HLA-C PCR reagents in the PCR set-up procedure so as to analyze the potential reasons for causing abnormal SBT result.</p><p><b>RESULTS</b>In the 620 samples typed at HLA-C locus using the AlleleSEQR HLA-C SBT commercial kit, 5 samples with no full match result were identified. The closest genotype showed one nucleotide mismatch with many different allele groups at different nucleotide position. Based on the PCR-SBT nucleotide sequence, heterozygous nucleotides were determined only in exon 4, whereas the nucleotides in exon 2 and 3 were all homozygotes. The results showed that HLA-Cw*0706 allele dropout existed in all the 5 samples with abnormal SBT results initially identified by AlleleSEQR HLA-C SBT kit, no novel mutation was found.</p><p><b>CONCLUSION</b>The results indicate that the PCR primer pair incompatible with DNA template may result in allele dropout in HLA-C SBT test. Based on the characterization of HLA-C full-length, it is essential to develop HLA-C SBT kit suitable for Chinese population in the future.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Reference Standards
/
Molecular Sequence Data
/
Base Sequence
/
HLA-C Antigens
/
DNA Fingerprinting
/
Amino Acid Sequence
/
Sequence Analysis, DNA
/
Alleles
/
Genetics
/
Methods
Limits:
Humans
Language:
Chinese
Journal:
Chinese Journal of Medical Genetics
Year:
2009
Type:
Article
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