Utility of a multiplex PCR assay for detecting herpesvirus DNA in clinical samples
Utility of a multiplex PCR assay for detecting herpesvirus DNA in clinical samples
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DOI：
10.1128/jcm.40.5.1728-1732.2002
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发表时间：
2002-05-01
影响因子：
9.4
通讯作者：
Birch, C
中科院分区：
文献类型：
--
作者：
Druce, J;Catton, M;Birch, C
多重PCR扩增单纯疱疹病毒1型和2型，巨细胞病毒，水痘带状疱疹病毒DNA存在于各种临床材料。这些病毒对至少一种抗病毒药物的体内抑制的敏感性是将其纳入多重检测系统的重要考虑因素。提取前，将等份马疱疹病毒引入每份标本中，作为PCR潜在抑制剂的指示剂和次优PCR条件的检测剂。与病毒分离和基于免疫荧光的抗原检测相比，多重检测对检测中代表的所有病毒的检出率更高。与用于鉴定这些病毒的其他技术相比，检测试剂盒的周转时间显著缩短。使用该检测方法进行了21，000多次检测。总的来说，多重PCR能够检测到大量增加的疱疹病毒，在某些情况下，在标本或解剖部位，以前很少，如果使用传统的检测方法确定。
A multiplex PCR was designed to amplify herpes simplex virus types 1 and 2, cytomegalovirus, and varicella-zoster virus DNA present in a diverse range of clinical material. The susceptibility of these viruses to in vivo inhibition by at least one antiviral drug was an important consideration in their inclusion in the multiplex detection system. An aliquot of equine herpesvirus was introduced into each specimen prior to extraction and served as an indicator of potential inhibitors of the PCR and a detector of suboptimal PCR conditions. Compared to virus isolation and immunofluorescence-based antigen detection, the multiplex assay yielded higher detection rates for all viruses represented in the assay. The turnaround time for performance of the assay was markedly reduced compared to those for the other techniques used to identify these viruses. More than 21,000 tests have been performed using the assay. Overall, the multiplex PCR enabled the detection of substantially increased numbers of herpesviruses, in some cases in specimens or anatomical sites where previously they were rarely if ever identified using traditional detection methods.