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Application of PCR-SSCP method for HLA DNA distinguish between homozygous and heterozygous genotype space ...

02,1301 = individualseachwithDPm0201, 0202,130] h. mogoteforSSCPon9acrylamidegels' discussion of PCR / SSO method is considered as the HLA of Microbiology and Immunology 15 September 1995 No. 5 stabbed DNA typing ~ kinds of simple, rapid and accurate method, almost all of the HLA used in laboratory. HLA-SSCP method can distinguish between homozygous and heterozygous genotypes blank genes. The results show: 9 samples brought 4 different DPB1 alleles, 12 or 9 in the polyacrylamide gel,[link widoczny dla zalogowanych], electrophoresis in the 22c 4 6 hours, can be effectively separated. and found that nine samples had a sample of four single-chain appears as the control of the hybrid zone and similar table. indicates that the sample of air from the hybrid gene. to consider the gap gene can not be sure that is not found in the new DPB1 alleles,[link widoczny dla zalogowanych], it could be 11IHWC (1991 years) and a few have been discovered after a new member of the DRBI alleles may also be false positive and false negative SSO hybridization of , in the type of information lost when DNA sequencing further confirmed depends. also found that heterozygous samples have also appeared with two bands of double-stranded, and the literature [s] reported that in line, but to distinguish between different heterozygotes with the double-stranded differences between the needs of a strong gel electrophoresis, so as SSCP simple and high resolution. 'in PcR / ssO classification, based on the locus in a sample of only one allele were detected, and further for PCR -SSCP analysis, no doubt to distinguish these samples were homozygous or heterozygous genotype is an effective method blank genes. Meanwhile, SSCP analysis is that there is no false positive, the detection rate as high as 8o ~ 9s, so get rid of SSO hybridization some false positive and false negative, is conducive to the discovery of new alleles. but simply using PCR-SSCP method for HLA typing, in a single block of gel to face a large number of alleles on the difficulties of distinguishing that the use of ARMS-SsCP (Ampliflea-tionrefractorymutations ~ temandsingle-stralldconformationpo [yi ~ orphism) HLA polymorphism analysis method is effective. But the pattern changes with a single chain complex and difficult to uniform classification standards. For SSCP method to replace PcR / ssO typing in HLADNA status, still need to develop and improve. Currently, the method as PcR / ssO a supplement, used to distinguish homozygous genes and genetic gaps, eliminate false positives and SSO hybridization of some false negative, for the improvement of DNA HLA typing is important to PCR / SSO methods for small samples forensic paternity testing and crime, it becomes time-consuming and costly and not practical. PCR-SSCP method is to overcome these shortcomings, and make it easy, fast and precise. Particular the use of non-radioactive silver staining method, to reduce the isotope contamination,[link widoczny dla zalogowanych], reduce costs, without reducing the sensitivity of silver staining, but often weak with a deep background of the display,[link widoczny dla zalogowanych], the method also continued to improve our findings, the work of the silver staining method used with the soaking solution with the new silver solution and the background color significantly lighter fluid can achieve good resolution results
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