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首页> 外文期刊>Journal of Clinical Microbiology >Combination of Reverse Transcriptase PCR Analysis and Immunoglobulin M Detection on Filter Paper Blood Samples Allows Diagnostic and Epidemiological Studies of Measles
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Combination of Reverse Transcriptase PCR Analysis and Immunoglobulin M Detection on Filter Paper Blood Samples Allows Diagnostic and Epidemiological Studies of Measles

机译：逆转录酶PCR分析与免疫球蛋白M检测相结合的滤纸血样可以对麻疹进行诊断和流行病学研究

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As measles control and elimination campaigns progress, laboratory confirmation of clinically diagnosed measles cases becomes increasingly important. However, in many tropical countries collection and storage of clinical specimens for this purpose are logistically complicated. In this study it is shown that blood samples spotted on filter paper are suitable for the laboratory diagnosis of measles using a combination of reverse transcriptase PCR (RT-PCR) analysis and immunoglobulin M (IgM) detection. First, it was shown that in vitro measles virus (MV)-infected cells diluted in human blood and spotted on filter paper can be detected by RT-PCR. Small amounts of infected cells remained detectable after 25 weeks of storage of the filter paper at room temperature, 4 weeks at 37°C, or 2 weeks at 45°C. Subsequently, this RT-PCR was applied to filter paper blood samples collected from 117 clinically diagnosed measles patients in Sudan in 1997 and 1998. Prior laboratory diagnosis had confirmed 90 cases as acute MV infections, while 27 proved to be nonmeasles rash disease cases. Positive RT-PCR signals were detected in filter paper blood samples of 43 of the 90 confirmed cases (48%) but in none of the 27 nonmeasles cases. In addition, MV-specific IgM levels measured in reconstituted filter paper samples correlated well with those measured in plasma samples. Measles diagnosis based on the combination of filter paper RT-PCR and IgM detection had a sensitivity and specificity of 99 and 96%, respectively. An advantage of this diagnostic approach is that sequencing of RT-PCR products allows phylogenetic analysis of the MV strain involved.
机译：随着麻疹控制和消除运动的进展，临床确诊的麻疹病例的实验室确认变得越来越重要。然而，在许多热带国家中，为此目的收集和存储临床标本在后勤方面很复杂。在这项研究中，通过结合逆转录酶PCR（RT-PCR）分析和免疫球蛋白M（IgM）检测，发现点样在滤纸上的血液样本适合于麻疹的实验室诊断。首先，已证明可以通过RT-PCR检测在人血中稀释并点在滤纸上的体外麻疹病毒（MV）感染细胞。在室温下将滤纸保存25周，37°C下4周或45°C下2周后，仍可检测到少量感染细胞。随后，该RT-PCR应用于从1997年和1998年在苏丹的117位临床诊断的麻疹患者中收集的滤纸血样。先前的实验室诊断已确认90例为急性MV感染，而27例被证实为非麻疹皮疹疾病病例。在90例确诊病例中的43例（48％）的滤纸血样中检测到了RT-PCR阳性信号，但在27例非麻疹病例中均未检测到。此外，在重构滤纸样品中测得的MV特异性IgM水平与血浆样品中测得的MV相关性很好。基于滤纸RT-PCR和IgM检测相结合的麻疹诊断的敏感性和特异性分别为99％和96％。这种诊断方法的优势在于，对RT-PCR产物进行测序可以对涉及的MV菌株进行系统发育分析。

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