SYNOPSIS



SYNOPSIS

Rajiv Gandhi University of Health Sciences, Karnataka, Bangalore.

“COMPARATIVE STUDY FOR DETECTION OF MYCOBACTERIA BY ACID FAST STAINING, LOWENSTEIN JENSEN MEDIUM AND MYCOBACTERIA GROWTH INDICATOR TUBE”

Name of the candidate : Dr. Pratibha Bhat. U

Guide : Dr. B. Rekha M.B.B.S, MD.,

Course and Subject : M.D. Microbiology.

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Department of Microbiology,

Father Muller Medical College,

Kankanady, Mangalore – 575002.

2010

RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES, KARNATAKA, BANGALORE.

ANNEXURE II

PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION

|1 |Name of the candidate and address (in block |Dr. PRATIBHA BHAT.U |

| |letters) |RESIDENT, |

| | |DEPARTMENT OF MICROBIOLOGY, |

| | |FATHER MULLER MEDICAL COLLEGE, |

| | |MANGALORE – 575002. |

|2 |Name of the Institution |FATHER MULLER MEDICAL COLLEGE MANGALORE. |

|3 |Course of study and Subject |M.D. Microbiology. |

|4 |Date of admission to course |30-04-2010 |

|5 |Title of the Topic |

| |COMPARATIVE STUDY FOR DETECTION OF MYCOBACTERIA BY ACID FAST STAINING, LOWENSTEIN JENSEN MEDIUM AND MYCOBACTERIA GROWTH INDICATOR|

| |TUBE. |

| | |

|6 |BRIEF RESUME OF THE INTENDED WORK: |

| |6.1 Need for the study |

| |Tuberculosis, one of the major airborne infectious bacterial diseases, remains a major worldwide health problem. The situation is |

| |further exacerbated with increasing incidence of drug resistant tuberculosis[1]. The advent of HIV infection, AIDS pandemic in 1980s|

| |has struck a blow on health care and there has been a global resurgence of tuberculosis. India is the highest tuberculosis burden |

| |country and accounts for nearly one fifth of global burden of tuberculosis[2]. |

| |The microscopic examination of sputum to detect presence of acid fast bacilli has served the world well for past 100 years and is |

| |diagnostic basis of directly observed therapy. Reliance solely upon the limited acid fast smear is one of the major factors that |

| |contribute to poor tuberculosis control in resource poor countries[3]. Bacteriological confirmation plays a key role in diagnosis of|

| |tuberculosis. The most commonly used conventional Lowenstein Jensen medium requires 6-10 weeks of incubation due to slow growth rate|

| |of Mycobacterium tuberculosis complex. The standard recommendation is that all new diagnostic systems to be used in screening |

| |patients are to be properly evaluated. It should be such that the laboratories must be able to isolate Mycobacteria easily, quickly |

| |and accurately [4]. |

| |Hence, there is need to evaluate newer methods for early detection of Mycobacterium tuberculosis complex, for effective treatment |

| |and control. |

| | |

| |6.2 Review of literature : |

| |Tuberculosis is responsible for about one third of preventable deaths worldwide[4]. The increasing incidence of tuberculosis and |

| |other mycobacterial diseases has made it essential for laboratories to quickly detect and identify Mycobacteria from human clinical |

| |material. |

| |Rodrigues et al[5] observed that in highly smear positive specimen (3+) a Mycobacteria Growth Indicator Tube culture scored almost |

| |99% recovery in 48-72 hours of culture inoculation and in case of smear negative specimen an average detection time was found to be |

| |16 days compared to 48 days by Lowenstein Jensen culture. Also, they found the system to be more advantageous in paucibacillary |

| |specimens like cerebrospinal fluid, body fluids and fine needle aspiration cytology samples. |

| |Dongsi Lu et al[6] showed in their study that although automated Mycobacteria Growth Indicator Tube system demonstrated better |

| |sensitivity than traditional Lowenstein Jensen slant for recovery of Mycobacteria from clinical specimens, approximately 10% of |

| |clinically significant Mycobacteria species would be missed by use of this system alone. Mycobacteria Growth Indicator Tube system |

| |is not yet sufficiently sensitive to warrant elimination of supplemental solid medium. |

| |Rishi et al[7] showed positivity rate of Mycobacteria Growth Indicator Tube960 system alone was 34.10% and of Lowenstein Jensen |

| |alone 1.93% and isolation rates of Mycobacteria in pulmonary and extra pulmonary samples were 61.83% and 21.98% by Mycobacteria |

| |Growth Indicator Tube, 44.01% and 4.96% by Lowenstein Jensen medium respectively. But for maximum recovery of Mycobacteria, a |

| |combination of Mycobacteria Growth Indicator Tube 960 and Lowenstein Jensen media should be used. |

| |6.3 Objectives of the study: |

| |1. To compare rate of detection of Mycobacteria by acid fast staining, Lowenstein Jensen medium and Mycobacteria Growth Indicator |

| |Tube culture methods. |

| |2. Characterization of Mycobacteria isolated. |

|7. |Material and methods: |

| |7.1 Source of data: About 100 patients suspected of pulmonary tuberculosis based on clinical history, attending Father Muller |

| |Medical College hospital will be studied. |

| |7.2 Method of collection |

| |Early morning expectorated sputum samples will be collected from clinically suspected cases of pulmonary tuberculosis in separate |

| |sterile, wide mouthed container. Quality of the sample will be assessed by macroscopic and microscopic examination. Acid fast |

| |staining will be done on the smear prepared from the specimen. The sample will be decontaminated by N-acetyl L-cysteine-sodium |

| |hydroxide (NALC-NaOH) method. This will be used to inoculate into Lowenstein Jensen medium and |

| | |

| |Mycobacteria Growth Indicator Tubes. The growth will be confirmed by acid fast staining and characterized by biochemical reactions. |

| | |

| |Selection Criteria: |

| |Inclusion Criteria: |

| |1. Patients with symptoms suggestive of pulmonary tuberculosis like- fever, cough with sputum for >2 weeks, night sweats, weight |

| |loss, anorexia, chest pain. |

| |2. Patients above 16 yrs of age. |

| |Exclusion Criteria: |

| |Patients currently on anti tubercular treatment. |

| |Extrapulmonary tuberculosis. |

| |Patients in paediatric age group. |

| |Data Analysis: |

| |The data collected will be analysed by Analysis of Variance (ANOVA) and Chi square test. |

| |7.3 Does the study require any investigations or interventions to be conducted on patients or other humans or animals? If so, please|

| |describe briefly. |

| |No |

| |7.4 Has ethical clearance been obtained from your institution? |

| |Yes |

|8 |LIST OF REFERENCES |

| |Negi SS, Khan SFB, Gupta S, Pasha ST et al. Comparison of conventional diagnostic modalities, BACTEC culture and polymerase chain |

| |reaction test for diagnosis of tuberculosis. Indian J Med Microbiol 2005;23(I):29-33. |

| |Park K, Epidemiology of Communicable Diseases Ch 5. In, Parks Textbook of Preventive and Social Medicine, M/S Banarsidas Bhanot |

| |Publishers, 20th edition, 2009;159-75. |

| |Espinal MA, Kim SJ, Suarez PG, Kam KM et al. Standard short course chemotherapy for drug resistant tuberculosis: treatment outcome in |

| |6 countries. JAMA 2000;283:2537-45. |

| |Chitra C and Prasad CE. Evaluation of Mycobacteria growth indicator tube (MGIT) for primary isolation of Mycobacteria. Ind.J Tub |

| |2001;48:155. |

| |Rodrigues C, Shenai S, Sadani M, Sukhadia N et al. Evaluation of BACTEC MGIT 960 TB system for recovery and identification of |

| |Mycobacterium tuberculosis complex in a high through put tertiary care center. Indian J Med Microbiol 2009;27(3):217-21. |

| |Dongsi Lu, Bobbi Heeren, Dunne Michael W. Comparison of the automated MGIT system with LJ medium for recovery of Mycobacteria from |

| |clinical specimens. Am J Clin Patho 2002;118:542-45. |

| |Rishi S, Sinha P, Malhotra B, Pal N. A comparative study for detection of Mycobacteria by BACTEC MGIT 960, LJ medium and direct AFB |

| |smear examination. Indian J Med Microbiol 2007;25(4):383-6. |

|9 |Signature of candidate | |

|10 |Remarks of the guide |Tuberculosis is one of the major infections seen in India, which requires to be |

| | |controlled. Early detection and prompt treatment is the need of the hour. This |

| | |study will help in comparing the newer methods with the existing methods in early |

| | |detection. |

|11 |Name & Designation of | |

| |(in block letters) | |

| |11.1 Guide |Dr. B. REKHA M.B.B.S, MD., |

| | |PROFESSOR AND HOD, |

| | |DEPT. OF MICROBIOLOGY, |

| | |FATHER MULLER MEDICAL COLLEGE, |

| | |KANKANADY, MANGALORE. |

| |11.2 Signature | |

| |11.5 Head of Department |DR. B. REKHA M.B.B.S, MD., |

| | |PROFESSOR AND HOD, |

| | |DEPARTMENT OF MICROBIOLOGY, |

| | |FATHER MULLER MEDICAL COLLEGE, |

| |11.6 Signature |KANKANADY, MANGALORE. |

|12 |12.1 Remarks of the | |

| |Chairman and Principal | |

| | | |

| | | |

| |12.2 Signature | |

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