School of Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney, NSW, Australia.
Zhejiang Provincial Center for Disease Control and Prevention, Hangzhou, China.
Front Cell Infect Microbiol. 2020 Sep 7;10:452. doi: 10.3389/fcimb.2020.00452. eCollection 2020.
spp. are a major cause of foodborne illness throughout the world. Traditional serotyping by antisera agglutination has been used as a standard identification method for many years but newer nucleic acid-based tests have become available that may provide advantages in workflow and test turnaround time. In this study, we evaluated the Luminex® xMAP® Serotyping Assay (SSA), a multiplex nucleic acid test capable of identifying 85% of the most common serotypes, in comparison to the traditional serum agglutination test (SAT) on 4 standard strains and 255 isolates from human (224), environmental, and food (31) samples. Of the total of 259 isolates, 256 could be typed by the SSA. Of these, 197 (77.0%) were fully typed and 59 (23.0%) were partially typed. By SAT, 246 of the 259 isolates (95%) were successfully typed. Sixty isolates had discrepant results between SAT and SSA and were resolved using whole genome sequencing (WGS). By SAT, 80.0% (48/60) of the isolates were consistent with WGS while by SSA 91.7% (55/60) were partially consistent with WGS. By serovar, all 30 serovars except one tested were fully or partially typable. The workflow comparison showed that SSA provided advantages over SAT with a hands-on time (HOT) of 3.5 min and total turnaround time (TAT) of 6 h, as compared to 1 h HOT and 2-6 days TAT for SAT. Overall, this study showed that molecular serotyping is promising as a rapid method for serotyping with good accuracy for typing most common serovars circulating in China.
spp. 是全世界食源性疾病的主要原因。多年来,传统的血清凝集试验一直被用作标准鉴定方法,但现已出现更新的基于核酸的检测方法,这些方法在工作流程和测试周转时间方面可能具有优势。在这项研究中,我们评估了 Luminex® xMAP® 血清分型检测试剂盒(SSA),这是一种能够鉴定 85%最常见血清型的多重核酸检测方法,与传统的血清凝集试验(SAT)在 4 株标准株和 255 株来自人类(224 株)、环境和食品(31 株)的分离株上进行了比较。在总共 259 株分离株中,256 株可通过 SSA 进行分型。其中,197 株(77.0%)完全分型,59 株(23.0%)部分分型。通过 SAT,259 株分离株中的 246 株(95%)成功分型。SAT 和 SSA 之间有 60 株分离株的结果存在差异,通过全基因组测序(WGS)解决了这些差异。通过 SAT,80.0%(48/60)的分离株与 WGS 一致,而通过 SSA,91.7%(55/60)与 WGS 部分一致。按血清型划分,除 1 个血清型外,其余 30 个血清型均可完全或部分分型。工作流程比较显示,SSA 的实际操作时间(HOT)为 3.5 分钟,总周转时间(TAT)为 6 小时,而 SAT 的 HOT 为 1 小时,TAT 为 2-6 天,因此 SSA 比 SAT 具有优势。总体而言,这项研究表明,分子血清分型作为一种快速的血清分型方法具有很大的应用前景,对于鉴定中国流行的大多数常见血清型具有良好的准确性。