Abebe A, Johansson B, Abens J, Strannegård O
Department of Virology, Karolinska Institute, Stockholm, Sweden.
Scand J Infect Dis. 1992;24(3):265-73. doi: 10.3109/00365549209061331.
A polymerase chain reaction (PCR) technique for the detection of human enteroviruses in stool specimens was developed. The test was based on the synthesis of cDNA, followed by PCR and slot blot hybridization. The primers used were selected from a highly conserved sequence in the 5'non-coding region of the enteroviral genome. By this method 27 different enterovirus serotypes (15 echo, 6 coxsackie A, 4 coxsackie B, poliovirus type 2 and enterovirus 71) from 89 patients could be detected. Using positive virus culture as reference, the sensitivity of PCR was 69% after 30 cycles of amplification, 91% using 30 + 10 cycles and 100% following 2 rounds of amplification with ensuing hybridization. None of 23 stool samples from healthy individuals or patients with meningitis of proven non-enteroviral etiology were positive by the PCR. By contrast, 13/26 culture-negative, randomly chosen stool samples from patients with suspected enteroviral disease were positive by the test. These findings demonstrate a high sensitivity and an apparently high specificity of PCR for detection of enteroviruses in stool samples. Therefore, the methodology may be useful in the laboratory diagnosis of enterovirus infections.
开发了一种用于检测粪便标本中人类肠道病毒的聚合酶链反应(PCR)技术。该检测基于cDNA的合成,随后进行PCR和狭缝印迹杂交。所用引物选自肠道病毒基因组5'非编码区的高度保守序列。通过这种方法,可以检测来自89名患者的27种不同肠道病毒血清型(15种埃可病毒、6种柯萨奇A病毒、4种柯萨奇B病毒、2型脊髓灰质炎病毒和71型肠道病毒)。以阳性病毒培养物作为参考,30个循环扩增后PCR的灵敏度为69%,30 + 10个循环时为91%,两轮扩增并随后杂交后为100%。23份来自健康个体或已证实非肠道病毒病因的脑膜炎患者的粪便样本经PCR检测均为阴性。相比之下,从疑似肠道病毒疾病患者中随机选取的26份培养阴性粪便样本中有13份经该检测呈阳性。这些结果表明PCR检测粪便样本中肠道病毒具有高灵敏度和明显的高特异性。因此,该方法可能有助于肠道病毒感染的实验室诊断。