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A Multiplex Nested PCR for the Simultaneous Detection of Salmonella typhi, Mycobacterium tuberculosis, and Burkholderia pseudomallei in Patients with Pyrexia of Unknown Origin (PUO) in Vellore, South India

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Abstract

Background and Objectives

Salmonella typhi, Mycobacterium tuberculosis, and Burkholderia pseudomallei are among the most important monocyte-tropic bacterial agents causing pyrexia of unknown origin (PUO), with a significant number of endemic infections in both South and Southeast Asian regions. These infections pose a major risk to travelers to these regions as well.

Methods

We developed and evaluated a multiplex nested polymerase chain reaction (PCR) for the simultaneous detection of the three pathogens in 305 patients’ buffy coat samples.

Results

The assay for S. typhi and B. pseudomallei was able to detect down to 1 colony forming unit/5 μL PCR input and M. tuberculosis was detected down to 20 genome copies/5 μL PCR input. S. typhi was detected in 10 (3.3 %) individuals, B. pseudomallei in 10 individuals (3.3 %), and M. tuberculosis in 18 individuals (5.9 %). Co-infections of M. tuberculosis and B. pseudomallei were detected in three individuals and S. typhi and B. pseudomallei in two individuals.

Conclusion

This protocol is efficient for PUO diagnosis especially in Asian countries.

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Acknowledgments and Disclosures

This research work was funded by the Indian Council of Medical Research (ICMR) grant no. 5/3/3/12/2010/ECD-I. The authors have no conflicts of interest that are directly relevant to the content of this article.

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Authors and Affiliations

  1. Sri Sakthi Amma Institute of Biomedical Research, Sri Narayani Hospital and Research Centre, Sripuram, Vellore, 632 055, Tamil Nadu, India

    Sathish Sankar, Kumaran Vadivel, Balaji Nandagopal, Mary V. Jesudason & Gopalan Sridharan

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  1. Sathish Sankar
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  2. Kumaran Vadivel
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  3. Balaji Nandagopal
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Correspondence to Sathish Sankar.

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Sankar, S., Vadivel, K., Nandagopal, B. et al. A Multiplex Nested PCR for the Simultaneous Detection of Salmonella typhi, Mycobacterium tuberculosis, and Burkholderia pseudomallei in Patients with Pyrexia of Unknown Origin (PUO) in Vellore, South India. Mol Diagn Ther 18, 315–321 (2014). https://doi.org/10.1007/s40291-013-0079-7

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