Viral Exanthemata and STD Section, National Microbiology Laboratory, Public Health Agency of Canada, JC Wilt Infectious Diseases Research Centre, 745 Logan Avenue, Winnipeg, Manitoba, Postal Code R3E 3L5, Canada.
Viral Exanthemata and STD Section, National Microbiology Laboratory, Public Health Agency of Canada, JC Wilt Infectious Diseases Research Centre, 745 Logan Avenue, Winnipeg, Manitoba, Postal Code R3E 3L5, Canada; Department of Medical Microbiology and Infectious Diseases, Faculty of Health Sciences, University of Manitoba, Winnipeg, Manitoba, Canada.
J Virol Methods. 2022 Jan;299:114349. doi: 10.1016/j.jviromet.2021.114349. Epub 2021 Nov 2.
Vaccination has greatly reduced global measles incidence, however measles remains endemic in many regions worldwide. Measles surveillance relies on high performance molecular detection of the virus. We have developed and validated a multiplex rRT-PCR assay for the detection of measles virus. The assay includes three independent probes with unique reporter dyes for the simultaneous detection of the measles hemagglutinin gene, nucleoprotein gene and endogenous RNaseP control. Using dilution series of synthetic RNAs the limits of detection were determined to be approximately 20 copies of measles RNA. The assay is extremely reproducible with very low intra-assay and inter-assay coefficients of varation for both the N and the H targets. After testing 68 confirmed measles positive and 86 measles negative archival clinical samples our data shows the multiplex assay has a sensitivity and specificity of 100 %, and a 100 % concordance with the expected results. No cross reactivity was identified with clinical specimens positive for six other viruses. According to the WHO, currently only the B3, D4, D8, H1 measles genotypes of the 24 recognized genotypes continue to circulate and this new multiplex assay successfully detected all four of those genotypes as well as six other genotypes.
疫苗接种大大降低了全球麻疹发病率，但麻疹在世界许多地区仍然流行。麻疹监测依赖于对病毒的高性能分子检测。我们开发并验证了一种用于检测麻疹病毒的多重实时 RT-PCR 检测方法。该检测方法包括三个具有独特报告染料的独立探针，用于同时检测麻疹血凝素基因、核蛋白基因和内源性 RNaseP 对照。使用合成 RNA 的稀释系列确定检测限约为 20 个麻疹 RNA 拷贝。该检测方法具有极高的重现性，N 和 H 靶标之间的内和间检测变异系数非常低。在测试了 68 个确诊的麻疹阳性和 86 个麻疹阴性存档临床样本后，我们的数据表明，多重检测方法的灵敏度和特异性均为 100%，与预期结果的符合率为 100%。与其他六种病毒的临床样本无交叉反应。根据世界卫生组织的说法，目前只有 24 种公认基因型中的 B3、D4、D8、H1 麻疹基因型继续传播，这种新的多重检测方法成功地检测到了所有四种基因型以及另外六种基因型。
