“FACTORS INFLUENCING POLYMERASE CHAIN

REACTION OUTCOMES IN PATIENTS

WITH CLINICALLY SUSPECTED
OCULAR TUBERCULOSIS”

Authors:
Praveen Kumar Balne, Rohit Ramesh Modi, Nuzhat
Choudhury, Neha Mohan, Manas Ranjan Barik,
Tapas Ranjan Padhi, Savitri Sharma, Satya Ranjan
Panigrahi and Soumyava Basu, 2014.

Presenter:
Manuel, Christine D.
 BACKGROUND OF THE STUDY

In paucibacillary infections such as ocular tuberculosis,

the polymerase chain reaction (PCR) assay can be used to
make a definitive diagnosis (TB). Infectious uveitis in
countries with a high endemicity. However, because this
disease is paucibacillary, conventional techniques such as
smear microscopy and culture are ineffective at detecting it.
NAATs, or nucleic acid amplification tests, are used to detect
mutations in DNA. As a result, polymerase chain reaction
(PCR) may be a viable option for determining the cause of
this condition. The paucibacillary nature of specimens, lack
of adequate sample volumes, and non-uniform distribution
make PCR-based diagnosis of any form of extrapulmonary TB
difficult. Bacteria in such specimens, the presence of PCR
inhibitors, and the lack of a proper gold standard for
assessing its diagnostic potential.
 OBJECTIV
E
To identify the different factors that
affect the polymerase chain reaction
outcomes in patient with clinically
suspected ocular TB.
 RESEARCH METHOD
PCR of aqueous or vitreous samples was performed on patients
with clinically suspected ocular TB. There were three control groups:
group 1 had culture-proven non-tuberculous endophthalmitis, group 2
had culture-negative non-tuberculous endophthalmitis, and group 3 had
uncomplicated cataract surgery. IS6110, MPB64, and protein b genes
from Mycobacterium tuberculosis complex were among the targets of
the PCR. Multiple regression analysis (5% level of significance) was
used to examine the relationships between positive PCR results and
disease laterality, tuberculin skin test (TST)/interferon-gamma release
assay (IGRA), chest radiography, and sample type (aqueous or
vitreous). Positive PCR by one or more gene targets, as well as factors
influencing positive PCR outcomes, were the primary outcome
measures.
 RESULT OF THE STUDY
MPB64, protein b, and IS6110 were all tested in all 114 samples. MPB64 was
detected in 70.2 % (n = 80) of the tested samples, protein B in 40.0 % (n = 44), and
IS6110 in only 9.1 % (n = 8). The DNA sequence of amplicons from four randomly
selected PCR reactions revealed M homology. Tuberculosis is a disease that affects the
entire body. Of the 80 PCR-positive patients, 71 completed a full course of
antitubercular therapy, with 65 (91.5 %) showing complete resolution of inflammation
at the final follow-up. 12.5 % (3 out of 24) of controls in group 1 and 18.7% (6 out of
32) of controls in group 2 tested positive by PCR. In control group 3 (n = 25), no
PCR-positive outcome was observed. Multiple regression analysis revealed a
significant relationship between a positive PCR outcome and bilateral presentation, but
not with a positive TST/IGRA, chest radiography, or sample type (aqueous/vitreous).