RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES, KARNATAKA

BANGALORE

ANNEXURE II
PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION

1. Name of the Candidate and address Dr. SUMALATHA V

POSTGRADUATE,
(in BLOCK letters) DEPARTMENT OF BIOCHEMISTRY,
BANGALORE MEDICAL COLLEGE &
RESEARCH INSTITUTE.

2. Name of the Institution BANGALORE MEDICAL COLLEGE&

RESEARCH INSTITUTE,
FORT,
BANGALORE-560 002.

3. Course of study and subject M.D. BIOCHEMISTRY

4. Date of admission to the course 08/06/2015

5. Title of Dissertation
A COMPARATIVE STUDY OF
INTERFERON GAMMA AND
ADA IN PLEURAL FLUID AS AN
EARLY MARKER OF
TUBERCULOUS PLEURAL
EFFUSION
6. BRIEF RESUME OF THE INTENDED WORK:

6.1 Need for the study:

Tuberculosis (TB) is the most common cause of pleural effusion in areas with
a high incidence of TB. The sensitivity of both direct microscopy and pleural
fluid cultures is relatively low, and the diagnosis is mainly based on the
presence of caseating granulomas in a pleural biopsy specimen. However, the
procedures for pleural tissue collection, such as blind pleural biopsy and
thoracoscopic biopsy are invasive procedures and are not without risk. Thus,
efforts to find alternative, rapid, noninvasive, and safe tests which maintain
high sensitivities and specificities for the diagnosis of pleural TB, have been
attempted. The measurement of adenosine deaminase (ADA) and interferon-
gamma (IFN-) in the supernatant of pleural fluid specimens may serve as one
example.1,2
6.2 Review of literature
1)Study done by Valdes et al,showed that ADA has diagnostic

value in Tuberculous pleural effusion1.

2)Study done by Aoe K et al,showed that IFN gamma in pleural

fluid is the most sensitive and specific marker of tuberculous

pleural effusion andand can be used as early markers2.

3)Study done by Villegas,etal,observed that ADA,PCR and

measurement of IFN gamma levels provide the basis for rapid

and efficient diagnosis of Pleural tuberculosis4.

4)Study done by Barnes PF,et al showed local production of TNF

And IFN gamma in tuberculous pleuritis5.

5)Study done by Ribera et al,observed that T Lymphocyte

produce high level of IFN gamma in tuberculous pleural

effusion6.
 6.3 Objectives of the study
1)Estimation of ADAand gamma IFNas a marker of tuberculosis.
2) To assess the correlation between gamma Interferonand ADA in
pleural fluid in the diagnosis of Tuberculosis.

MATERIALS AND METHODS

7.1 Source of data :The study will comprise of in-patients diagnosed to

have with pleural effusion from the department of Medicine of Victoria
Hospital and Bowring and Lady Curzon Hospitals, Bangalore

7.2 Methods of collection of data :

A. Study design:Observational,prospective,case control study.
B. Study period : November 2015-May 2017
7.
C. Place of study :Victoria Hospital,Bowring and Lady Curzon
Hospital

D. Sample size :After consulting the statistician,30 cases and 30 controls of

pleural effusion patients are taken from Victoria ,Bowring and Lady Curzon
Hospital.

E. Inclusion criteria:

1. Patients who give written informed consent.

 2. Patients between the age group of 20- 60 years admitted with

exudative pleural effusion.

3. Non tuberculosis pleural effusion(malignancy, transudate)

individuals as controls.

F. Exclusion criteria

a.Patients with pleural infection due to viral infection.

b. Pregnancy and lactating females.
c. Patients with cutaneous drug reactions.
d. Patients with empyema.

G. Methodology :

Inpatients diagnosed with pleural effusion from the department of Pulmonary

Medicine of Victoria Hospital and Bowring and Lady Curzon Hospitals,
Bangalore and fulfilling the inclusion and exclusion criteria will be taken into
study.

History and clinical examination will be recorded in the study

proforma.Patients will undergothoracocentesis in the first 24 hours after the
admission. Pleural fluid was analyzed for biochemical markers, Gram stain,
and bacterial and TB culture, as well as for cytology and differential white
blood cell count using standard cytospin procedures and hematoxylin-eosin or
Papanicolaou stains. Effusions were classified as transudates or exudates,
using Lights criteria.

Pleural fluid samples (mean volume 50 mL) for ADA and IFN-
 measurements were centrifuged at 2000 revolutions per minute for 10 minutes,
and the supernatant was frozen at 80C until assayed for markers.

H. Assessment tools:

a. Proforma for written informed patient consent.(Annexure-1).

b. Studyproforma. (Annexure-2).

I. Statistical analysis

Suitable statistical method will be used at the time of analysis.

7.3 Does the study require any investigations, interventions to be

conducted on patients or other humans or animals? If so please describe
briefly.

YES
The study requires the following investigations to be conducted on patients and
controls.

Parameters to be studied
1. Pleural fluid Interferon gamma levels.
2. Pleural fluid Adenosine Deaminase levels
 3.Cell count and cell type
4.Pleural fluid analysis for sugar,protein
5.X-Ray thorax( P-A view)

7.4 Has ethical clearance been obtained from your institution in case of
7.3?

YES.

LIST OF REFERENCES

(1.)Valds L, Alvarez D, San Jos E, et al. Value of adenosine deaminase in the diagnosis of
8.
tuberculous pleural effusions in young patients in a region of high prevalence of
tuberculosis. Thorax. 1995;50(6):600603.
(2.) Aoe K, Hiraki A, Murakami T, et al. Diagnostic significance of interferon-gamma in
tuberculous pleural effusions. Chest. 2003;123(3):740744.
(3.)Crompton GK, McHardy GF. Diseases of the respiratory system. In: Edwards CR, Boucher

IA. Davidson's Principles and Practice of Medicine. 16 th ed. Edinburgh:ELBS;1992.p.410-2.

(4.)Villegas MV, Labrada LA, Saravia NG. Evaluation of polymerase chain reaction, adenosine
deaminase, and interferon-gamma in pleural fluid for the differential diagnosis of pleural
tuberculosis. Chest 2000;118:1355-64
(5.)Barnes PF, Fong SJ, Brennan PJ, Twomey PE, Mazumder A, Modlin RL. Local production of
tumor necrosis factor and interferon gamma in tuberculouspleuritis. J Immunol 1990;145:149-54.

(6.) Ribera E, Ocaa I, Martinez-Vazquez JM,Rossell M, Espaol T, Ruibal A. High level of

interferon gamma in tuberculous pleural effusion. Chest 1998;93:30811

(7.) Sderblom T, Nyberg P, Teppo AM, Klockars M, RiskaH,Pettersson T. Pleural fluid interferon-
gamma and tumour necrosis factor-alpha in tuberculous and rheumatoid pleurisy. EurRespir J
1996;9:1652-5.

(8.) Light RW. Disorders of pleura, mediastinum and diaphragm. In:Braunwald E, Fauci AS, Kasper
DL, Hauser SL, Longo DL, Jameson JL.Harrison's Principles of Internal Medicine, vol.2., 15
thed.New York: McGraw Hill International Edition; 2003:1513-5.

(9.) Villena V, Lpez-Encuentra A, Echave-Sustaeta J, Martin-Escribano P, Ortuo-de-Solo

B,Estenoz-Alfaro J. Interferon-gamma in 388 immunocompromised and immunocompetent patients
for diagnosing pleural tuberculosis. EurRespir J 1996;9:2635-9.
9. Signature of the candidate:
10. Remarks of the guide : Analysis of pleural fluid interferon gamma
has more specificity and sensitivity than
pleural fluid ADA levels in diagnosis of
tuberculosis.

11. Name and Designation of Dr. B.V. MARUTHI PRASADM.D

Guide:( in block letters) PROFESSOR OF BIOCHEMISTRY,,
11.1 Guide: DEPARTMENT OF BIOCHEMISTRY,
BANGALORE MEDICAL COLLEGE AND
RESEARCH INSTITUTE, BANGALORE.

11.2Signature :

11.3Co-Guide: Dr. C. NAGARAJA M.D

PROFESSOR AND HEAD ,
DEPARTMENT OF PULMONARY
MEDICINE,
VICTORIA HOSPITAL,
BANGALORE MEDICAL COLLEGE AND
RESEARCH INSTITUTE, BANGALORE.

11.4 Signature:

11.5 Head of the Department: Dr. H. L. VISHWANATH M.D.

PROFESSOR AND HEAD,
DEPARTMENT OF BIOCHEMISTRY,
BANGALORE MEDICAL COLLEGE AND
RESEARCH INSTITUTE, BANGALORE.

11.6Signature:
12. 12.1 Remarks of the chairman
and Principal:

12.2 Signature: