基于PCR的流感病毒感染诊断或可通过唾液实现

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免疫层析抗原检测技术曾被广泛用于流感病毒的检测；然而其低灵敏度限制了该技术只能使用鼻咽拭子作为测试材料。唾液可以无创地获得，并对流感的诊断是有效的。聚合酶链式反应（PCR）的耗时性使得其不常用于快速诊断。

近日，来自日本信州大学医院的科学家采集了144例患者的鼻咽拭子和唾液，采用反转录定量聚合酶链反应（RT-PCR）和微滴式RT-PCR进行研究，评价了基于PCR分析的、以唾液为流感病毒感染诊断材料的检测方法。结果发表在《临床化学学报》（Clinica Chimica Acta，CCA）。

结果显示，鼻咽拭子和唾液两者结果的整体一致性达95.8%。使用微滴式RT-PCR技术可以在12分钟内检测到唾液中的流感病毒基因。每微升唾液以及鼻咽拭子含有超过1 × 102拷贝的流感病毒基因。在流感症状出现的24小时内，将近一半的患者的唾液和鼻咽拭子结果均呈阳性。

本研究表明，唾液可以作为鼻咽拭子的替代生物样本。当小型化机械实现快速PCR分析和RNA提取的全自动化以后，基于PCR的及时检测可以通过使用无材料限制的唾液而实现。

来源：艾兰博曼医学网

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Evaluation of saliva as diagnostic materials for influenza virus infection by PCR-based assays

Akane Suekia, Kazuyuki Matsudaa, , , Akemi Yamaguchib, Masayuki Ueharab, Mitsutoshi Suganoa, Takeshi Ueharaa, Takayuki Hondaa

Abstract

Background

Immunochromatographic antigen tests have been widely used for detection of influenza virus; however its low sensitivity restricts the use of clinical materials other than nasopharyngeal swabs. Saliva is obtained non-invasively and has utility for diagnosis of influenza. Polymerase chain reaction (PCR) is not typically used for rapid testing because it is time consuming. We evaluated the utility of saliva as diagnostic materials for influenza virus infection by PCR-based assays.

Methods

Nasopharyngeal swabs and saliva were simultaneously collected from 144 patients and investigated by reverse transcription-quantitative PCR (RT-qPCR) and droplet-RT-PCR.

Results

Overall concordance of results from nasopharyngeal swabs and saliva were 95.8%. Influenza gene was detectable in less than 12 min in saliva by the droplet-RT-PCR. Saliva as well as nasopharyngeal swabs contained more than 1 × 102 copies/μl of the influenza gene. About half of the patients provided positive results in nasopharyngeal swabs and saliva within 24 h from the onset of the symptoms.

Conclusion

The study demonstrates that saliva can be used as an alternative specimen source to nasopharyngeal swabs. When rapid PCR assay including RNA extraction to be full-automation in a miniaturized machine, point-of-care test based on PCR may be realized using saliva without restriction of materials.

http://www.sciencedirect.com/sci ... i/S0009898115300759