Sueki Akane, Matsuda Kazuyuki, Yamaguchi Akemi, Uehara Masayuki, Sugano Mitsutoshi, Uehara Takeshi, Honda Takayuki
Department of Laboratory Medicine, Shinshu University Hospital, Matsumoto, Japan.
Department of Laboratory Medicine, Shinshu University Hospital, Matsumoto, Japan.
Clin Chim Acta. 2016 Jan 30;453:71-4. doi: 10.1016/j.cca.2015.12.006. Epub 2015 Dec 4.
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.
Nasopharyngeal swabs and saliva were simultaneously collected from 144 patients and investigated by reverse transcription-quantitative PCR (RT-qPCR) and droplet-RT-PCR.
Overall concordance of results from nasopharyngeal swabs and saliva were 95.8%. Influenza gene was detectable in less than 12min in saliva by the droplet-RT-PCR. Saliva as well as nasopharyngeal swabs contained more than 1×10(2) copies/μl of the influenza gene. About half of the patients provided positive results in nasopharyngeal swabs and saliva within 24h from the onset of the symptoms.
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.
免疫层析抗原检测已广泛用于流感病毒检测;然而,其低灵敏度限制了除鼻咽拭子以外的临床材料的使用。唾液采集无创,对流感诊断有实用价值。聚合酶链反应(PCR)通常不用于快速检测,因为它耗时。我们通过基于PCR的检测方法评估了唾液作为流感病毒感染诊断材料的效用。
从144例患者中同时采集鼻咽拭子和唾液,并通过逆转录定量PCR(RT-qPCR)和液滴RT-PCR进行检测。
鼻咽拭子和唾液检测结果的总体一致性为95.8%。通过液滴RT-PCR在唾液中不到12分钟即可检测到流感基因。唾液和鼻咽拭子中流感基因的含量均超过1×10(2)拷贝/μl。约一半的患者在症状出现后24小时内鼻咽拭子和唾液检测结果呈阳性。
该研究表明唾液可作为鼻咽拭子的替代标本来源。当包括RNA提取在内的快速PCR检测在小型仪器中实现全自动化时,基于PCR的即时检测可能无需材料限制即可使用唾液实现。