Non-specificity of sequence characterised amplified region as an alternative molecular epidemiology marker for the identification of Salmonella enterica subspecies enterica serovar Typhi

Ja’Afar Nuhu Ja’Afar,Kia Kien Phua,Subhash Janardhan Bhore

doi:10.1186/s13104-018-3870-z

Abstract

ObjectiveIdentification of Salmonella Typhi by conventional culture techniques is labour-intensive, time consuming, and lack sensitivity and specificity unlike high-throughput epidemiological markers that are highly specific but are not affordable for low-resource settings. SCAR, obtained from RAPD technique, is an affordable, reliable and reproducible method for developing genetic markers. Hence, this study investigated the use of SCAR as an alternative molecular epidemiological marker for easy identification of S. Typhi in low-resource settings.ResultsOne hundred and twenty RAPD primers were screened through RAPD-PCR against a panel of common enterobacteriaceae for the best RAPD band pattern discrimination to develop SCAR primers that were used to develop a RAPD-SCAR PCR. Of this number, 10 were selected based on their calculated indices of discrimination. Four RAPD primers, SBSA02, SBSA03, SBSD08 and SBSD11 produced suitable bands ranging from 900 to 2500 bp. However, only SBSD11 was found to be specific for S. Typhi, and was cloned, sequenced and used to design new SCAR primers. The primers were used to amplify a panel of organisms to evaluate its specificity. However, the amplified regions were similar to other non-Typhi genomes denoting a lack of specificity of the primers as a marker for S. Typhi.

Highlights

The amplified regions were similar to other non-Typhi genomes denoting a lack of specificity of the primers as a marker for S
Typhoid fever, a systemic disease caused by the bacterium Salmonella enterica subspecies enterica serovar Typhi
Typhi isolates (STY083 (ATCC 7251), STY088 and STY231) that had been previously differentiated by pulsed-field gel electrophoresis (PFGE)

Summary

Introduction

A systemic disease caused by the bacterium Salmonella enterica subspecies enterica serovar Typhi Typhi), is global in distribution but more prevalent in Oceania, Africa, Latin America and Asia with prevalence rates of 15.4, 49.8, 53.1, and 274.3 per 100,000 population, respectively [1]. Conventional culture techniques are labour intensive, time consuming, expensive, and lack sensitivity and specificity [2, 3]. Typhi isolates are often so similar and look identical using most typing techniques [4]. High throughput epidemiological markers such as pulse-field gel electrophoresis (PFGE) and single nucleotide polymorphism (SNP) markers are employed to track and monitor S. Typhi and the disease it causes [7–9]. These markers are expensive to develop and are not readily affordable in low-resource settings where the disease is mostly endemic

Results

Discussion

Conclusion