Multiplex PCR assay for simultaneous detection and differentiation of Entamoeba histolytica, Giardia lamblia, and Salmonella spp. in the municipality-supplied drinking water

Prem Shankar; Jyotsna Mishra; Vijaya Bharti; Deepak Parashar; Sarman Singh

doi:10.4103/JLP.JLP_66_18

Journal of Laboratory Physicians, Table of Contents

CC BY-NC-ND 4.0 · J Lab Physicians 2019; 11(03): 275-280
DOI: 10.4103/JLP.JLP_66_18

Original Article

Multiplex PCR assay for simultaneous detection and differentiation of Entamoeba histolytica, Giardia lamblia, and Salmonella spp. in the municipality-supplied drinking water

Prem Shankar

Department of Laboratory Medicine, Division of Clinical Microbiology and Molecular Medicine, All India Institute of Medical Sciences, New Delhi, India

,

Jyotsna Mishra

Department of Anesthesiology, Medical College of Wisconsin, Milwaukee, Wisconsin, USA

,

Vijaya Bharti

Department of Pharmacology, Vanderbilt University, Nashville, Tennessee, USA

,

Deepak Parashar

Department of Obstetrics and Gynecology, Medical College of Wisconsin, Milwaukee, Wisconsin, USA

,

Sarman Singh

Department of Laboratory Medicine, Division of Clinical Microbiology and Molecular Medicine, All India Institute of Medical Sciences, New Delhi, India

› Author Affiliations

Abstract

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Abstract

BACKGROUND: The contamination with Entamoeba histolytica, Giardia lamblia, and Salmonella spp. in drinking water is the most prevalent in Indian subcontinent, but often difficult to detect all these pathogens from the drinking water.

MATERIALS AND METHODS: A multiplex polymerase chain reaction (mPCR) method was developed to detect contamination of municipality-supplied drinking water with E. histolytica, G. lamblia, and Salmonella spp. The primers were designed to target small subunit of 16S rRNA type gene of E. histolytica and G. lamblia, and invasive A gene of Salmonella typhimurium. The optimized mPCR assay was applied on 158 municipality-supplied drinking water samples collected from Delhi.

RESULTS: Out of total 158 water samples, 89 (56.32%) were found positive for the targeted pathogens by mPCR while conventional methods could be detected only in 11 (6.96%) samples. The mPCR assay showed 100% sensitivity and specificity for these pathogens in comparison with culture and microscopic detection. Of the 89 mPCR-positive samples, G. lamblia, E. histolytica, and Salmonella spp. were present in 35 (22.15%), 26 (16.45%), and 28 (17.72%), respectively. Nine (5.69%) samples were positive for both E. histolytica and G. lamblia, 10 (6.32%) were positive for G. lamblia and Salmonella spp., and 8 (5.06%) had Salmonella spp. and E. histolytica. Nonetheless, 3 (1.89%) samples were positive for all three pathogens.

CONCLUSIONS: The present assay is an alternative to conventional methods to serve as highly sensitive, specific, and economical means for water quality surveillance to detect the outbreak caused by E. histolytica, G. lamblia, and Salmonella spp. pathogens.

Key words

Drinking water - Entamoeba histolytica - Giardia lamblia - multiplex polymerase chain reaction - Salmonella species

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References

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