title: 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.
abstract: OBJECTIVE
  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.
  
  
  CONCLUSIONS
  This protocol is efficient for PUO diagnosis especially in Asian countries.
authors.affiliation: Sri Sakthi Amma Institute of Biomedical Research, Sri Narayani Hospital and Research Centre, Sripuram, Vellore, 632 055, Tamil Nadu, India, sathish3107@gmail.com.
authors.forename: Sathish
authors.initials: S
authors.lastname: Sankar
authors.forename: Kumaran
authors.initials: K
authors.lastname: Vadivel
authors.forename: Balaji
authors.initials: B
authors.lastname: Nandagopal
authors.forename: Mary V
authors.initials: MV
authors.lastname: Jesudason
authors.forename: Gopalan
authors.initials: G
authors.lastname: Sridharan
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001424:Bacterial Infections / Q000382:microbiology / Q000473:pathology*; D016957:Burkholderia pseudomallei / Q000235:genetics / Q000302:isolation & purification*; D002648:Child; D002675:Child, Preschool; D004269:DNA, Bacterial / Q000235:genetics; D005260:Female; D005335:Fever of Unknown Origin / Q000382:microbiology*; D006801:Humans; D007194:India; D007223:Infant; D008297:Male; D008554:Melioidosis / Q000382:microbiology / Q000473:pathology; D008875:Middle Aged; D009169:Mycobacterium tuberculosis / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012485:Salmonella typhi / Q000235:genetics / Q000302:isolation & purification*; D014376:Tuberculosis / Q000382:microbiology / Q000473:pathology; D014435:Typhoid Fever / Q000382:microbiology / Q000473:pathology; D055815:Young Adult
chemical_list: D004269:DNA, Bacterial