Evaluation of Mycobacterium tuberculosis 16 and 71 Heat Shock Proteins in the Cerebrospinal Fluid of Tuberculous Meningitis Patients
Seema D. Shekhawat
Biochemistry Research Laboratory, Central India Institute of Medical Sciences, Nagpur - 440 010, India
Rajpal S. Kashyap *
Biochemistry Research Laboratory, Central India Institute of Medical Sciences, Nagpur - 440 010, India
Snehal S. Wanjari
Biochemistry Research Laboratory, Central India Institute of Medical Sciences, Nagpur - 440 010, India
Khushboo J. Nagdev
Biochemistry Research Laboratory, Central India Institute of Medical Sciences, Nagpur - 440 010, India.
Hemant J. Purohit
Environmental Genomics Unit, National Environmental Engineering Research Institute, Nehru Marg, Nagpur-440020, India
Girdhar M. Taori
Biochemistry Research Laboratory, Central India Institute of Medical Sciences, Nagpur - 440 010, India
Hatim F. Daginawala
Biochemistry Research Laboratory, Central India Institute of Medical Sciences, Nagpur - 440 010, India
*Author to whom correspondence should be addressed.
Abstract
Aims: To evaluate the efficiency of two potential Mycobacterium tuberculosis (M. tb) heat shock proteins (Hsps) towards the improvement of tuberculous meningitis (TBM) diagnosis.
Study Design: The patients were divided into TBM (confirmed and suspected) and non TBM group. The cerebrospinal fluid (CSF) was collected and evaluated for M. tb Hsp 16 and 71.The Indirect ELISA results of M. tb Hsp 71 were compared with polymerase chain reaction (PCR).
Place and Duration of Study: Biochemistry Research Laboratory, Central India Institute of Medical Sciences, Nagpur between June 2009 and July 2010.
Methodology: 29 TBM and 22 non TBM CSF samples were collected. Indirect ELISA was performed for evaluating the, M. tb Hsp16 and Hsp71 in the collected samples, individually as well as in combination. The ELISA method for detection of M. tb Hsp 71 was also compared with in house PCR technique for TBM diagnosis.
Results: The data analysis was done with MedCalc® Software. M. tb Hsp16 showed positivity of 58.62% and negativity of 68.18%. Similarly for M. tb Hsp71, positivity is 89.65% and negativity is 68.18%.The results of ELISA for M. tb Hsp71 was compared with PCR technique and concordance was also calculated. Of the Hsp ELISA positive group for M. tb Hsp71, 24 were PCR positive and 2 were PCR negative with the 92.30 % concordance in TBM patients and in non TBM patients the concordance was observed to be 93.30%. Use of the monoclonal antibody Hsp 71 appear preferable over individual use of M. tb Hsp 16 and combined use of both Hsp and yield optimum results.
Conclusion: Our data suggest that the detection of M. tb Hsp 71 in the CSF sample of TBM patients can be useful for the diagnosis of TBM patients. These predictors, however, need further work to validate reliability.
Keywords: Tuberculous meningitis, heat shock proteins, ELISA, chaperones