Line Probe Assay, The Best Molecular Diagnostic Tool and Improve the Multi Drug Resistance Detection in Pulmonary and Extra Pulmonary Tuberculosis Cases in Balochistan

Abstract

Introduction:

Mycobacterium tuberculosis (MTB) appears resistant to first line antibiotics e.g. rifampicin and isoniazid is said to be Multi-drug resistant Tuberculosis(MDR-TB), Globally 15.3 million cases emerge where as in Pakistan 1.8 million cases reported included Pakistan in 22 high burden cases. For MDR-TB detection in this study LPA a molecular technique was used.

Methods: A number 400 patients were studied in this study after sample collection in open space for pulmonary sample and for extra pulmonary samples were taken in sterile syringe or containers auramine staining was performed and examine under 40X lenes, DNA Extinction ; the DNA was extracted by Genolyse Method, Master Mix preparation: The master mix was adjusted by mixing of master mix A and mix B for further uses. e.g. Amplification and hybridization: for amplification process thermocycler was used as PCR amplification and for hybridization the PCR products were analyzed  to the strips on which probe are presents and interpreted

Results: The study was conducted among different ethnic groups and both male & female genders of Balochistan. Overall 400 smear negative and positive samples from TB DOTS and PMDTS were applied on LPA for the diagnosis of Mono and MDR-TB. In total 400 cases, 236 (59%) were MTB positive, 162 (40%) were found MTB negative and 2 were detected as invalid. In this study also analysis were based on gender(male and Female), ethnicity (Pashtoon, Baloch, Hazara and Sattler’s )and site (DOTS and PMDTS).

Conclusion: in conclusion MDR-TB detection by LPA is more sensitive technique to others and prevalence rate of MDR-TB in enrolled patients at PMDTS and DOTS program at alarming stage, a serious threats  to public. to overcome this issue molecular LPA is must for the MTB diagnosis on routine bases and to make possible reduction of MDR-TB cases. The outcomes of this study enhance about the mutation occurs in MTB is highly which promote TB in MDR-TB and in future MDR-TB in to XDR-TB. It is highly suggested for the development in the diagnostic techniques and tools .i.e.LPA  for MDR-TB detection.