Hsia K, Spector D H, Lawrie J, Spector S A
Department of Pediatrics, University of California, San Diego, La Jolla 92093.
J Clin Microbiol. 1989 Aug;27(8):1802-9. doi: 10.1128/jcm.27.8.1802-1809.1989.
Polymerase chain reaction (PCR) amplification was used to detect human cytomegalovirus (HCMV) sequences. The fragments selected for amplification were fragments of 130 and 152 base pairs (bp) located at two opposite ends of HCMV strain AD169 EcoRI fragment D. Amplification of the 152-bp DNA was consistently greater than that of the 130-bp DNA. At the optimal Mg2+ concentration of 5 mM, specific PCR amplification of 152-bp DNA with Taq polymerase was sensitive; only one AD169-infected fibroblast cell or 0.01 pg of AD169 fragment D DNA was needed for detection. This specific amplification was also found with various clinical HCMV isolates and peripheral blood cells and urine from patients. In 37 urine samples analyzed simultaneously by PCR and by virus cultivation, identical results were found in 35 samples, while 2 scored positive only by PCR. This suggests that specific amplification of 152-bp DNA is sensitive and can be used for rapid detection of HCMV infections.
采用聚合酶链反应（PCR）扩增法检测人巨细胞病毒（HCMV）序列。选择用于扩增的片段是位于HCMV AD169株EcoRI片段D两端的130和152个碱基对（bp）的片段。152 bp DNA的扩增始终大于130 bp DNA的扩增。在最佳Mg2+浓度为5 mM时，用Taq聚合酶对152 bp DNA进行特异性PCR扩增很灵敏；检测仅需一个感染AD169的成纤维细胞或0.01 pg的AD169片段D DNA。在各种临床HCMV分离株以及患者的外周血细胞和尿液中也发现了这种特异性扩增。在同时通过PCR和病毒培养分析的37份尿液样本中，35份样本结果相同，而2份样本仅PCR检测呈阳性。这表明152 bp DNA的特异性扩增很灵敏，可用于快速检测HCMV感染。
