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。
