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Rapid and Sensitive Detection of Major Uropathogens in a Single-Pot Multiplex PCR Assay

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Abstract

Urinary tract infection (UTI) is among the most common bacterial infections and poses a significant healthcare burden. Escherichia coli is the most common cause of UTI accounting for up to 70 % and a variable contribution from Proteus mirabilis, Pseudomonas aeruginosa and Klebsiella pneumoniae. To establish a complete diagnostic system, we have developed a single-tube multiplex PCR assay (mPCR) for the detection of the above-mentioned four major uropathogens. The sensitivity of the assay was found to be as low as 102 cfu/ml of cells. The mPCR evaluated on 280 clinical isolates detected 100 % of E. coli, P. aeruginosa, P. mirabilis and 95 % of K. pneumonia. The assay was performed on 50 urine samples and found to be specific and sensitive for clinical diagnosis. In addition, the mPCR was also validated on spiked urine samples using 40 clinical isolates to demonstrate its application under different strain used in this assay. In total, mPCR reported here is a rapid and simple screening tool that can compete with conventional biochemical-based screening assays that may require 2–3 days for detection.

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Acknowledgments

We acknowledge Dr Sumathi, Government General Hospital Chennai, for providing some of the samples used in this study.

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Author notes

  1. B. M. Jaffar Ali

    Present address: Centre for Green Energy Technology, Pondicherry University, Puducherry, 605 014, India

Authors and Affiliations

  1. AU-KBC Research Centre, M.I.T. Campus of Anna University, Chennai, 600 044, India

    B. Padmavathy, R. Vinoth Kumar, Amee Patel, S. Deepika Swarnam & B. M. Jaffar Ali

  2. Sundaram Medical Foundation and Dr. Rangarajan Memorial Hospital, Chennai, 600 040, India

    T. Vaidehi

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  1. B. Padmavathy
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  2. R. Vinoth Kumar
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Correspondence to B. M. Jaffar Ali.

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Padmavathy, B., Vinoth Kumar, R., Patel, A. et al. Rapid and Sensitive Detection of Major Uropathogens in a Single-Pot Multiplex PCR Assay. Curr Microbiol 65, 44–53 (2012). https://doi.org/10.1007/s00284-012-0126-3

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  • DOI: https://doi.org/10.1007/s00284-012-0126-3

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