Du Tian, Lin Ji-Hong, Zhao Jun-Hua, Wang Hai-Bo, Mo Qiu-Hua
Futian District Center for Disease Control and Prevention, Shenzhen 518040, Guangdong, China.
Zhongshan International Travel Healthcare Center, Zhongshan 528400, Guangdong, China.
Can J Infect Dis Med Microbiol. 2020 Sep 7;2020:9373984. doi: 10.1155/2020/9373984. eCollection 2020.
and are often associated with fecal-oral transmission and cause large-scale outbreaks in centralized catering units and, therefore, should be frequently and strictly monitored, especially among food handlers. However, no specific and sensitive on-site detection method is available until now.
In this study, an insulated isothermal PCR assay for the detection of and on a field-deployable PCR system was developed. Specificity, sensitivity, reproducibility, and clinical accuracy of the assay were characterized and evaluated.
The insulated isothermal PCR assay could be completed within 58 minutes with minimal pretreatment needed. The assay was specific and with good reproducibility. The limit of detection was 10 CFU/mL and 10 CFU/mL for and , respectively, which was comparable to multiplex real-time PCR. Mock on-site clinical evaluation results showed that the analytical sensitivity and specificity of the insulated isothermal PCR assay were 100% and 96.6%, while the positive predictive value and negative predictive value were 94.1% and 100%, respectively.
Based on our results, we believe that the assay developed herein could serve as an alternative method for preliminary screening and provide a valuable platform for the on-site detection of and , especially in resource-limited and developing countries.
[具体病原体1]和[具体病原体2]通常与粪口传播相关，并在集中供餐单位引发大规模疫情，因此，应经常且严格地进行监测，尤其是在食品从业人员中。然而，目前尚无特异性强且灵敏的现场检测方法。
在本研究中，开发了一种用于在现场可部署的PCR系统上检测[具体病原体1]和[具体病原体2]的隔热等温PCR检测方法。对该检测方法的特异性、灵敏度、可重复性和临床准确性进行了表征和评估。
隔热等温PCR检测方法在只需最少预处理的情况下可在58分钟内完成。该检测方法具有特异性且重复性良好。[具体病原体1]和[具体病原体2]的检测限分别为10 CFU/mL和10 CFU/mL，与多重实时PCR相当。模拟现场临床评估结果显示，隔热等温PCR检测方法的分析灵敏度和特异性分别为100%和96.6%，而阳性预测值和阴性预测值分别为94.1%和100%。
基于我们的结果，我们认为本文开发的检测方法可作为一种初步筛查的替代方法，并为[具体病原体1]和[具体病原体2]的现场检测提供一个有价值的平台，特别是在资源有限的发展中国家。
