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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

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...

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Published in:Molecular diagnosis & therapy 2014-06, Vol.18 (3), p.315-321
Main Authors: Sankar, Sathish, Vadivel, Kumaran, Nandagopal, Balaji, Jesudason, Mary V., Sridharan, Gopalan
Format: Article
Language:English
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Summary: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.
ISSN:1177-1062
1179-2000
DOI:10.1007/s40291-013-0079-7