应用 PCR 技术鉴定与马里发热患者侵袭性感染相关的非伤寒沙门氏菌血清型。

Identification by PCR of non-typhoidal Salmonella enterica serovars associated with invasive infections among febrile patients in Mali.

机构信息

Center for Vaccine Development, University of Maryland School of Medicine, Baltimore, MD, USA.

出版信息

PLoS Negl Trop Dis. 2010 Mar 9;4(3):e621. doi: 10.1371/journal.pntd.0000621.

BACKGROUND

In sub-Saharan Africa, non-typhoidal Salmonella (NTS) are emerging as a prominent cause of invasive disease (bacteremia and focal infections such as meningitis) in infants and young children. Importantly, including data from Mali, three serovars, Salmonella enterica serovar Typhimurium, Salmonella Enteritidis and Salmonella Dublin, account for the majority of non-typhoidal Salmonella isolated from these patients.

METHODS

We have extended a previously developed series of polymerase chain reactions (PCRs) based on O serogrouping and H typing to identify Salmonella Typhimurium and variants (mostly I 4,[5],12:i:-), Salmonella Enteritidis and Salmonella Dublin. We also designed primers to detect Salmonella Stanleyville, a serovar found in West Africa. Another PCR was used to differentiate diphasic Salmonella Typhimurium and monophasic Salmonella Typhimurium from other O serogroup B, H:i serovars. We used these PCRs to blind-test 327 Salmonella serogroup B and D isolates that were obtained from the blood cultures of febrile patients in Bamako, Mali.

PRINCIPAL FINDINGS

We have shown that when used in conjunction with our previously described O-serogrouping PCR, our PCRs are 100% sensitive and specific in identifying Salmonella Typhimurium and variants, Salmonella Enteritidis, Salmonella Dublin and Salmonella Stanleyville. When we attempted to differentiate 171 Salmonella Typhimurium (I 4,[ 5],12:i:1,2) strains from 52 monophasic Salmonella Typhimurium (I 4,[5],12:i:-) strains, we were able to correctly identify 170 of the Salmonella Typhimurium and 51 of the Salmonella I 4,[5],12:i:- strains.

CONCLUSION

We have described a simple yet effective PCR method to support surveillance of the incidence of invasive disease caused by NTS in developing countries.

背景

在撒哈拉以南非洲地区，非伤寒沙门氏菌（NTS）已成为婴幼儿侵袭性疾病（菌血症和局灶性感染，如脑膜炎）的主要病因。重要的是，包括来自马里的数据在内，三种血清型——肠炎沙门氏菌血清型 Typhimurium、肠炎沙门氏菌血清型 Enteritidis 和都柏林沙门氏菌——导致了从这些患者中分离出的大多数非伤寒沙门氏菌。

方法

我们扩展了先前开发的一系列基于 O 群和 H 型的聚合酶链反应（PCR），以鉴定肠炎沙门氏菌血清型 Typhimurium 和变异体（主要是 I 4,[5],12:i:-）、肠炎沙门氏菌血清型 Enteritidis 和都柏林沙门氏菌。我们还设计了引物来检测在西非发现的沙门氏菌 Stanleyville 血清型。另一个 PCR 用于区分两相肠炎沙门氏菌血清型 Typhimurium 和单相肠炎沙门氏菌血清型 Typhimurium 与其他 O 群 B、H:i 血清型。我们使用这些 PCR 对来自马里巴马科发热患者血液培养物中分离的 327 株 B 群和 D 群沙门氏菌血清型进行盲法检测。

主要发现

我们已经证明，当与我们之前描述的 O 群 PCR 联合使用时，我们的 PCR 在识别肠炎沙门氏菌血清型 Typhimurium 和变异体、肠炎沙门氏菌血清型 Enteritidis、都柏林沙门氏菌和沙门氏菌 Stanleyville 方面具有 100%的灵敏度和特异性。当我们试图区分 171 株肠炎沙门氏菌血清型 Typhimurium（I 4,[5],12:i:1,2）和 52 株单相肠炎沙门氏菌血清型 Typhimurium（I 4,[5],12:i:-）菌株时，我们能够正确识别出 170 株肠炎沙门氏菌血清型 Typhimurium 和 51 株沙门氏菌 I 4,[5],12:i:- 菌株。