title: Factors influencing polymerase chain reaction outcomes in patients with clinically suspected ocular tuberculosis.
abstract: BACKGROUND
  Polymerase chain reaction (PCR) assay can be a useful method for definitive diagnosis in paucibacillary infections such as ocular tuberculosis (TB). In this study, we have evaluated factors affecting PCR outcomes in patients with clinically suspected ocular TB. Patients with clinically suspected ocular TB were investigated by PCR of aqueous or vitreous samples. Three control groups were also tested: group 1 included culture-proven non-tuberculous endophthalmitis, group 2 culture-negative non-tuberculous endophthalmitis, and group 3 patients undergoing surgery for uncomplicated cataract. PCR targeted one or more of following targets: IS6110, MPB64, and protein b genes of Mycobacterium tuberculosis complex. Multiple regression analysis (5% level of significance) was done to evaluate the associations between positive PCR outcome and laterality of disease, tuberculin skin test (TST)/interferon-gamma release assay (IGRA), chest radiography, and type of sample (aqueous or vitreous). The main outcome measures were positive PCR by one or more gene targets, and factors influencing positive PCR outcomes.
  
  
  RESULTS
  All 114 samples were tested for MPB64, 110 for protein b, and 88 for IS6110. MPB64 was positive in 70.2% (n = 80) of tested samples, protein b in 40.0% (n = 44), and IS6110 in only 9.1% (n = 8). DNA sequencing of amplicons from four randomly chosen PCR reactions showed homology for M. tuberculosis complex. Of the 80 PCR-positive patients, 71 completed a full course of antitubercular therapy, of which 65 patients (91.5%) had complete resolution of inflammation at final follow-up. Among controls, 12.5% (3 out of 24) in group 1 and 18.7% (6 out of 32) in group 2 also tested positive by PCR. No PCR-positive outcome was observed in control group 3 (n = 25). Multiple regression analysis revealed significant association of positive PCR outcome with bilateral presentation, but not with a positive TST/IGRA, chest radiography, or type of sample (aqueous/vitreous) used.
  
  
  CONCLUSIONS
  Careful selection of gene targets can yield high PCR positivity in clinically suspected ocular TB. Bilateral disease presentation but not any evidence of latent systemic TB influences PCR outcomes. False-positive results may be seen in ocular inflammation unrelated to ocular TB.
authors.forename: Praveen Kumar
authors.initials: PK
authors.lastname: Balne
authors.forename: Rohit Ramesh
authors.initials: RR
authors.lastname: Modi
authors.forename: Nuzhat
authors.initials: N
authors.lastname: Choudhury
authors.forename: Neha
authors.initials: N
authors.lastname: Mohan
authors.forename: Manas Ranjan
authors.initials: MR
authors.lastname: Barik
authors.forename: Tapas Ranjan
authors.initials: TR
authors.lastname: Padhi
authors.forename: Savitri
authors.initials: S
authors.lastname: Sharma
authors.forename: Satya Ranjan
authors.initials: SR
authors.lastname: Panigrahi
authors.affiliation: L V Prasad Eye Institute, Patia, Bhubaneswar, Odisha 751 024, India. eyetalk@gmail.com.
authors.forename: Soumyava
authors.initials: S
authors.lastname: Basu