Lynas C, Hurlock N J, Copplestone J A, Prentice A G, McGonigle R J
Department of Haematology, Derriford Hospital, Plymouth.
J Clin Pathol. 1994 Jul;47(7):609-12. doi: 10.1136/jcp.47.7.609.
To determine whether the polymerase chain reaction with sequence specific primers (PCR-SSP) can assign HLA-DR type more accurately than serology in a routine hospital laboratory.
The 93 patients currently awaiting kidney transplants have been DR typed by serology over the past 14 years, 82% within the past five years. They have now been retyped using the PCR-SSP method described by Bein et al. Where the two results differed, PCR-SSP was repeated, once by the same method and once using the primer set of Olerup and Zetterquist.
There were 13 (14%) discrepancies between the results. Of these, two were PCR-SSP failures, later overcome: three were failure to detect DRB1*0103 by serology; five assignment of other alleles by PCR-SSP to serological "blanks"; and three alleles were differently assigned by serology and PCR. The serological typing of the final patient when repeated for this study was at variance with the original findings (14 years ago), but in agreement with PCR. In the remaining patients, serology had not determined the split of 36 DR3 alleles (all DR17 by PCR-SSP) or 13 DR6 alleles (12 DR13 and one DR14 by PCR-SSP). One patient in each case had their antigen splits of DR2 and DR5 assigned by PCR-SSP (DR15 and DR11, respectively) but not by serology.
PCR-SSP provides more reliable and detailed information on HLA-DR polymorphism than serology, and does so within a routine tissue typing laboratory.
确定在医院常规实验室中,序列特异性引物聚合酶链反应(PCR-SSP)在HLA-DR分型上是否比血清学方法更准确。
过去14年中,93例等待肾移植的患者已通过血清学进行DR分型,其中82%是在过去5年内进行的。现在使用Bein等人描述的PCR-SSP方法对他们进行重新分型。当两种结果不同时,重复进行PCR-SSP,一次使用相同方法,一次使用Olerup和Zetterquist的引物组。
结果之间存在13例(14%)差异。其中,2例PCR-SSP失败,后来得以克服;3例血清学未能检测到DRB1*0103;5例PCR-SSP将其他等位基因指定为血清学“空白”;3个等位基因在血清学和PCR分型中结果不同。在本研究中,对最后一名患者的血清学分型结果进行重复检测时,与最初结果(14年前)不一致,但与PCR结果一致。在其余患者中,血清学未确定36个DR3等位基因(PCR-SSP均为DR17)或13个DR6等位基因(PCR-SSP为12个DR13和1个DR14)的细分情况。每种情况下各有1例患者的DR2和DR5抗原细分情况通过PCR-SSP确定(分别为DR15和DR11),但血清学未能确定。
在常规组织分型实验室中,PCR-SSP在HLA-DR多态性方面比血清学提供更可靠、更详细的信息。