Browsing by Author "Kaewkungwal, J"
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Publication katG (SER 315 THR) Gene Mutation in Isoniazid Resistant Mycobacterium tuberculosis(Kathmandu University, 2011) Marahatta, SB; Gautam, S; Dhital, S; Pote, N; Jha, AK; Mahato, R; Mishra, S; Poudel, BH; Ramasoota, P; Kaewkungwal, J; Singhasivanon, PABSTRACT Background Isoniazid (INH) together with Rifampicin (RFP) forms the cornerstone of a short chemotherapy course for tuberculosis (TB) treatment. Mutation at codon 315 of katG gene is most prevalent in isoniazid resistant Mycobacterium tuberculosis (MTB) and is high in area with high TB incidence. Polymerase Chain Reaction Restriction Fragment Length Polymorphism (PCR-RFLP) has been found to be a reliable and effective tool for the identification of the specific gene alteration. Objective The objective of this study was to screen Ser315Thr mutation of KatG gene of INH resistant MTB strain by PCR-RFLP technique. Methods Altogether 37 INHr MTB isolates obtained from German Nepal Tuberculosis Project (GENETUP) Kathmandu Nepal was included in the study. Deoxyribonucleic Acid (DNA) extraction was performed according to protocol of SORPOCLEAN™ from the culture isolates. Amplification of the fragment with katG codon 315 was performed in a Biometra Thermocycler using primers. The amplified fragment was cleaved with MspI. The restriction fragments obtained were electrophoresed in a 2% agarose gel and were visualized using transilluminator. Results The katG Ser315Thr mutation was observed in 23 (62.2%) out of 37 INH resistant isolates. The drug susceptibility profile of INHr MTB isolates showed all isolates to be resistant to INH and RFP whereas 26 and 27 MTB isolates were resistant to Ethambutol (EMB) and Streptomycin (S) respectively. Seventeen (17) patients were harbouring katG gene mutated strain among Ethambutol and Streptomycin resistant cases. Conclusion The study identified high prevalence of Ser315Thr mutation in katG. The isolates harbouring this mutation were also simultaneously resistant to RFP. Ser315Th could be a potential genetic marker for predicting MDR-TB. KEY WORDS Isoniazid resistant MTB, katG gene, Mycobacterium tuberculosis, PCR-RFLP, mutation.Publication Risk factors of Multidrug Resistant Tuberculosis in central Nepal: A pilot study(Kathmandu University, 2010) Marahatta, SB; Kaewkungwal, J; Ramasoota, P; Singhasivanon, PABSTRACT Introduction Tuberculosis is the most widespread infectious disease in Nepal and poses a serious threat to the health and development of the country. Incidences of drug resistant tuberculosis in Nepal are increasing and this tuberculosisis a major threat to successfully controlling tuberculosis . Objective The general objective of the study was to assess the risk factors of multi-drug resistant tuberculosis among the patients attending the National Tuberculosis Centre, Bhaktpur Nepal. Methods An observational study/ case-control study with a Atotal number of 55 multi-drug resistant tuberculosis cases and 55 controls. The study was conducted among the patient attending in the National Tuberculosis Centre , Bhaktpur Nepal for six months, between May–October 2010. sImulti-drug resistant tuberculosis wasThe collected data was analysed in SPSS 11.5 version. The association between categorical variables were analysed by chi-square tests, OR and their 95% CI were measured. Results The total number of patients used for the study was 110, of which among them 55 were cases and 55 were controls . Our study revealed that there were significant associations between history of prior TB MDR-TB OR =2.799 (95 % CI 1.159 to 6.667) (p=0.020); smoking habit OR =2.350 and (95%CI 1.071 to 5.159) (p=0.032); social stigma social stigma OR 2.655 (95%CI r 1.071 to 5.159) (p=0.013); knowledge on MDR-TB OR =9.643 (95% CI 3.339 to 27.846) (p < 0.001)and knowledge on DOTS Plus OR=16.714 (95% CI is ranging from 4.656 to 60.008) (p< 0.001). However, there was no association found between alcohol drinking habits and ventilation in the room. Conclusion Our study revealed that there were significant associations between history of prior tuberculosis, smoking habit social stigma social stigma, knowledge on multi- drug resistant tuberculosis and knowledge on DOTS Plus with multi-drug resistant tuberculosis However there was no association between alcohol drinking habit and ventilation in room with multi-drug resistant tuberculosis. Key Words directly observed treatment short course-plus, multidrug-resistant tuberculosis, risk factors