Browsing by Subject "Mycobacterium tuberculosis"
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Item Comparative Study of Growh of mycobacterium Tuberculosis on Modified Ogawa Media, Modified Lowenstein Jensen Media and 2% Buffered Lowenstein Jensen (BLJ) Media(Department of Microbiology, 2008) Amatya, Sajeen BahadurThis prospective hospital based study was conducted from December 2005 to September 2006 atGerman-Nepal Tuberculosis Project- National Reference Laboratory (GENETUP-NRL),Kalimati, Kathmandu. In the present study, a total of 324 sputum samples from suspected TBpatients were collected. In all the samples 220 were smear positive and 104 were smear negativesamples as determined microscopically by standard fluorochrome method. In all the samples 177 were collected from patients suspected of pulmonary tuberculosis. All ofthe 324 sputum samples were divided into two equal halves and transferred in two calibratedcentrifuge tubes. All of the samples were decontaminated by 4% NaOH (Petroff’s method) andNekal-BX methods separately. Each decontaminated sample was cultured on three differentculture media; viz Modified Lowenstein-Jensen (MLJ), Modified Ogawa(MOG) and newlyproposed 2% Buffered Lowenstein-Jensen (BLJ) media. In all the smears positive samples 94.091% samples was culture positive in one or all culturemedia used. In all smear negative samples 9.62% samples were culture positive at least in oneculture media. Among smear positive samples decontaminated by 4% NaOH, 91.36%, 81.36%and 92.27% samples were culture positive on MOG, MLJ and BLJ medium, respectively.Similarly, in all smear positive samples decontaminated by Nekal, 92.72% were culturepositivein MOG, 87.73% in MLJ and 92.72% in BLJ. In all the smear negative samples decontaminated by 4% NaOH, 3.85%, 2.88% and 3.85%samples were culture positive on MOG, MLJ and BLJ medium. Similarly, among smear negativesamples decontaminated by Nekal, 4.81% were culture positive in MOG, 5.77% in MLJ and3.85% in BLJ. The MOG yielded more positive result (92.05%) followed by MLJ (84.32%) (P<0.01) and BLJ(92.50%). Among the decontamination techniques the Nekal method yielded better result(87.27%) than 4% NaOH (81.3%) for the recovery of bacteria on MLJ (P<0.05). No significantdifference obtained among samples decontaminated by 4% NaOH (91.36%) and Nekal (92.73%)with respect to recovery of bacteria on MOG (P>0.05). Similarly, no significant difference obtained among samples decontaminated by 4% NaOH(92.27%) and Nekal (92.73%) for the recovery of bacteria on BLJ (P>0.05). In all the smearnegative samples decontaminated by Nekal, highest culture positive result was observed on MLJ(5.77%) followed by BLJ (5.77%). While no significant difference was observed in the case of Nekal or 4% NaOH decontaminationmethod with respect to recovery of bacteria on MOG and BLJ. The result indicated that the BLJ medium is suitable for the recovery of bacteria followed byMOG and MLJ. However, the addition of 2% monopotassium phosphate buffer in MLJ yieldedsignificant increase in culture positive result. Similarly, the Nekal method was suitable decontamination method than 4% NaOH, for the better recovery of bacteria on MLJ.Bacteriological examination of sputum is essential for the diagnosis of pulmonary tuberculosis.The detection of acid-fast bacilli (AFB) in smears of sputum sample by Ziehl-Neelsen staining provides fastest evidence of the presence of Mycobacteria. However, the definitive diagnosis of Tuberculosis demands sputum positive culture for the M. tuberculosis which depends on the suitable decontamination method of the sputum sample and selection of suitable culture media. Key words:Mycobacterium tuberculosis, Decontamination and Homogenization, Petroff’s 4% NaOH method, Nekal-BX, Modified Lowenstein-Jensen media (MLJ), Modified Ogawa media(MOG), 2% Buffered Lowenstein-Jensen media (BLJ).Item Evaluation of Nitrate Reductase Assay For Rapiddetection of drugresistanttuberculosis(Central Department of Microbiology, 2010) Sah, Ranjit KumarDue to the emergence of multidrug-resistant tuberculosis (MDR-TB), there is an urgentdemand for simple, rapid and inexpensive methods of detecting drug resistanttuberculosisfor effectivetreatment, particularly in low-income countries.This study hasthe objective to evaluate nitrate reductase assay (NRA) efficacy for streptomycin,isoniazid, rifampicin and ethambutol susceptibility testing of Mycobacteriumtuberculosis strains.This prospective study was carried out at National TBReferenceLaboratory and SAARC TB and HIV/AIDS Centre, Thimi, Bhaktapur, Nepal fromNovember 2009 to May 2010.A total of113 clinical isolates ofM. tuberculosisweretested for four first line antitubercular drugs by nitrate reductase assay and werecompared with standard proportion method. The sensitivity and specificity of NRAwere98.1% and 100% for isoniazid, 95.1% and 98.6% for rifampicin,91.4% and 94.9%for streptomycin, and 78.6% and 97.9% for ethambutol respectively. AgreementbetweenNRA and proportion method were99.1%, 97.3%, 93.8%, 95.6%for isoniazid,rifampicin, streptomycin and ethambutol respectively.Results were available in 7-14days by NRAas compared to proportion method which takes 4-6 wk.NRA is reliablefor susceptibility testing of isoniazid and rifampicin, the two most important drugs forthe treatment of tuberculosis and is also easy to perform and inexpensive. In addition,the reduction in the timenecessary to obtain susceptibility results is of fundamentalimportance. Key words:Drug susceptibility,Mycobacterium tuberculosis,nitrate reductase assay,Proportion methodItem Extrapulmonary Tuberculosis of patient Visiting Tribhuvan University Teaching Hospital (TUTH) Kathmandu, Nepal(Department of Microbiology, 2010) Pudasaini, MeeraTuberculosis is one of the giant killer disease which accounts for massive death allover the world. Extra pulmonary tuberculosis becomes a serious problem through out the globe including Nepal.The samples were collected and decontaminated and AFB staining as well as culture on Ogawa medium was done. The isolates were identifiedon the basis of biochemical tests.Within the study period from June 2006 to Feb 2007, a total of 150 samples were collected from the patients visited to the Tribhuvan University, Teaching Hospital (TUTH) suspected of Extrapulmonary Tuberculosis. A varieties of Extra pulmonary specimens were brought to the Mycobacteriology laboratory, that includes Urine, Endometrial biopsy, Pleuralfluid, peritoneal fluid, Cerebrospinal fluid, Blood, As citicfluid and Pus. The samples were collected , decontaminated , AFB stained and cultured on Ogawa medium. The isolates were identified on the basis of biochemical tests.Of the total 150 samples, 40 were from male patients and 110 were from female patients.The highest percentage of suspected patients (54.67%) was found in the age group 20-30 followed by 30-40 years. The highest number of female patients 67(60.91%) were found in the age group 21-30 years followed by 12 (10.91%) in 30-40years. Similarly, the highest number of male patients 15 (37.50%) were found in the age group 20-30 years followed by 10 (25.00%) in 30-40 years.The highest number of samples 65 (43.33%) was endometrial biopsy followed by urine 30 (20%). The least number of samples were blood 2 (1.33%).Out of 150 cases of suspected extrapulmonary T.B, only 18 (12%) cases were found to be positive. Among 150 samples, 10 (6.67%) showed positive result from direct smear, 13(8.67%), showed positive result from culture and only 5 (3.33%) were found to be positive from both direct smear and culture.The highest percentage of positive cases9 (49.99%) was found in the age group 20-30 followed by 5 (27.77%) in 10-20 years.The highest number of female patients7 (38.88%) were found in the age group 20-30years followed by 2 (11.11%) in 10-20 years. Similarly, the highest number of male patients 3 (16.66%) were found in the age group 10-20 years followed by 2 (11.11%)in 20-30 years.The highest number of positive cases in both the direct smear examination and culture was obtained from pus sample followed by pleural fluid and urine samples.All the isolated culture tested for Niacin and Nitrate reduction test were found to be positive and negative for heat labile catalase test. Thus, the isolates were confirmed as Mycobacterium tuberculosis.Key Words:Extra pulmonary tuberculosis, Ogawa medium, direct smear, Culture, Mycobacterium tuberculosis.Item Prevalance of Pulmonary Tuberculosis in Patients Visiting National Tuberculosis Centre(Department of Zoology, 2021) Husain, AshikTuberculosis is a main public health problem all over the world. On an average one third of the world’s population is infected with Mycobacterium tuberculosis and problem of being in a state of existence disease. Tuberculosis remains a major threat in developing country like Nepal. Most of the cases occur in south-east Asia and Africa. This study mainly aims to find prevalence in terms of age, sex and to assess the knowledge tuberculosis. The study was analytical and hospital based conducted from December 2019 to February 2020 in National Tuberculosis Centre in Bhaktapur. During the study period a total of 765 sputum samples were collected and examined in the laboratory of National Tuberculosis Centre using fluorescence microscopy to detect the Mycobacterium and to assess the knowledge a total of 100 suspected patients were interviewed with closed ended questionnaire with multiple responses. Among the suspected patients, 73 (7%) showed positive with acid fast bacillus (AFB) and males (62%) were found to be more infected than female (38%) which was statistically insignificant (χ xii 2 =0.903, P> 0.05). The highest prevalence was found in age groups 51-60 (22.64%) which was statistically significant (F=39.6, P<0.05) between ages. There is mixed response showing good knowledge in affected parts and symptoms with statistically significant association (F=12.594, P<0.05) in the views of male and female. In general there is inadequate knowledge in terms of causative agent, preventive measures and treatment duration. Effective strategies must be implemented to bring equality to seek health behavior in males and females are required. Strategy with major focus on cultural and social bias must be integrated to bring more people aware of the disease.Item Retrospective Assessment of the Trend of Tuberculosis (TB) at Lumbini Provincial Hospital During 2015 to 2020 along with Knowledge Attitude and Practice among TB Patients(Department of Zoology, 2021) K.C, ShreestiPulmonary tuberculosis (TB) is an infectious bacterial disease infecting the lungs caused by Mycobacterium tuberculosis. The morbidity and mortality due to TB is decreasing in trend in most of the countries. To assess the disease trend particularly in Lumbini province as well as people's current perception of the disease, a retrospective analysis of the disease along with the Knowledge, attitude and practices (KAP) survey was carried out at Lumbini Provincial Hospital (LPH). Five years of data analysis of the Lumbini Provincial Hospital from 2015 AD indicated that the disease condition was in a steady state with slight increase (9.5%) in 2016/17. Suspected people seeking a diagnosis of TB in the hospital seems continuously increased from 2015 to 2020. TB prevalence for the last 5 years indicated 7.25% using AFB smear microscopy and 36.47% by Xpert MTB/RIF method. Males were found highly infected compared to female and mostly infected age group were 15-30 years. Similarly based on caste group Madeshi and Janjati people were found highly infected compared to others. Questionnaire survey results indicated that awareness of the disease is high but the knowledge of symptoms and route of the disease transmission is unsatisfactory. For preventive practices of the disease that people followed seems poor. Hence, the disease condition is still worsening the life of the people in the province. To reduce the disease burden, community-focused awareness and preventive program are recommended.