Orle K A, Gates C A, Martin D H, Body B A, Weiss J B
Roche Molecular Systems, Alameda, California 94501, USA.
J Clin Microbiol. 1996 Jan;34(1):49-54. doi: 10.1128/jcm.34.1.49-54.1996.
A multiplex PCR (M-PCR) assay with colorimetric detection was devised for the simultaneous amplification of DNA targets from Haemophilus ducreyi, Treponema pallidum, and herpes simplex virus (HSV) types 1 and 2. By using target-specific oligonucleotides in a microwell format, 298 genital ulcer swab specimens collected in New Orleans during three intervals from 1992 through 1994 were evaluated. The results of the M-PCR assay were compared with the results of dark-field microscopy and H. ducreyi culture on two different culture media. HSV culture results were available for 99 specimens collected during the third interval. Confirmatory PCR assays targeting different gene sequences for each of the three organisms were used to validate the M-PCR results. Specimens were resolved as positive for the determination of sensitivity if the reference diagnostic test was positive or if the results of both the M-PCR and the confirmatory PCR were positive. The resolved sensitivities of M-PCR for HSV, H. ducreyi, and T. pallidum were 100, 98.4, and 91%, respectively. The resolved sensitivities of HSV culture, H. ducreyi culture, and dark-field microscopy were 71.8, 74.2, and 81%, respectively. These results indicate that the M-PCR assay is more sensitive than standard diagnostic tests for the detection of HSV, H. ducreyi, and T. pallidum from genital ulcers.
设计了一种采用比色检测的多重聚合酶链反应（M-PCR）检测方法，用于同时扩增来自杜克雷嗜血杆菌、梅毒螺旋体以及1型和2型单纯疱疹病毒（HSV）的DNA靶标。通过在微孔板中使用靶标特异性寡核苷酸，对1992年至1994年三个时间段在新奥尔良收集的298份生殖器溃疡拭子标本进行了评估。将M-PCR检测结果与暗视野显微镜检查结果以及在两种不同培养基上进行的杜克雷嗜血杆菌培养结果进行了比较。对于在第三个时间段收集的99份标本，可获得HSV培养结果。针对这三种生物体各自不同基因序列的验证性PCR检测用于验证M-PCR结果。如果参考诊断测试呈阳性，或者M-PCR和验证性检测结果均为阳性，则将标本判定为检测敏感性阳性。M-PCR对HSV、杜克雷嗜血杆菌和梅毒螺旋体的检测敏感性分别为100%、98.4%和91%。HSV培养、杜克雷嗜血杆菌培养和暗视野显微镜检查的检测敏感性分别为71.8%、74.2%和81%。这些结果表明，M-PCR检测方法在从生殖器溃疡中检测HSV、杜克雷嗜血杆菌和梅毒螺旋体方面比标准诊断测试更敏感。
