SYNOPSIS

RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES,

KARNATAKA, BANGALORE

“COMPARITIVE STUDY ON THIN SMEAR, THICK SMEAR, QBC METHOD IN THE LABORATORY DIAGNOSIS OF MALARIA”

Name of the candidate : MR. SUJITH JOHN

Guide : DR.HILDA FERNANDES

Course and Subject : M.Sc MEDICAL LABORATORY TECHNOLOGY

( HAEMATOLOGY AND BLOOD TRANSFUSION )

DEPARTMENT OF PATHOLOGY

FR. MULLER MEDICAL COLLEGE HOSPITAL,

KANKANADY, MANGALORE – 575 002

RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES

KARNATAKA, BANGALORE

ANNEXURE –II

PROFORMA FOR REGISTRATION OF SUBJECTS FOR PROJECTS

1 NAME OF THE CANDIDATE AND ADRESS

Mr. SUJITH JOHN

M.Sc. MEDICAL LABORATORY

TECHNOLOGY

(HAEMATOLOGY & BLOOD TRANSFUSION)

FATHER MULLER MEDICAL COLLEGE,

MANGALORE-575002

2 NAME OF THE INSTITUTION FATHER MULLER MEDICAL COLLEGE

KANKANADY, MANGALORE- 575002

3 COURSE OF STUDY AND SUBJECT

MSc. MEDICAL LABORATORY

TECHNOLOGY

(HAEMATOLOGY & BLOOD TRANSFUSION)

4 DATE OF ADMISSION TO COURSE

1-10-12

5 TITLE OF THE TOPIC:

“COMPARATIVE STUDY ON THIN SMEAR, THICK SMEAR, QBC METHOD IN THE LABORATORY DIAGNOSIS OF MALARIA ”

6 BRIEF RESUME OF THE INTENDED WORK

6.1 NEED FOR THE STUDY:

Malaria causes 1.5 to 2.7 million deaths each year world wide1. The earliest

symptoms of malaria are very non specific and variable . So the early diagnosis of malaria

is very necessary inorder to avoid complications2. Therefore precise laboratory diagnosis

and species identification is essential3.

Microscopic examination of thick and thin blood smear is the standard method for

malaria diagnosis. The method is easily available and has low cost but its reliability is

questionable at low level parasitaemia4. This is labour intensive and depends on the quality

of the stained preparation as well as of microscopes, besides requiring considerable skill

for its interpretation5.

The QBC (Quantitative buffy coat) technique is a recently developed, quick technique

for malaria diagnosis. The staining property of malarial parasite with fluroscent dyes like

acridine orange, which is not depicted by the RBC host and the reduced buoyant density of

the infected erythrocytes are the two properties that are exploited in QBC technique for

malaria diagnosis6.

The present study aims at comparing the ability of QBC ,thick smear, thin smear in

the detection of malarial parasite.

6.2 REVIEW OF LITERATURE

Malaria is one of the most successful parasite ever known to mankind. After

thousands of years, it remains the world's most pervasive infection, affecting at least 91

different countries and some 300 million people. The disease causes fever, shivering, joint

pain, headache, and vomiting. In severe cases, patients can have jaundice, kidney failure,

and anaemia, and can lapse into a coma . It is ever-present in the tropics and countries in

sub-Saharan Africa, which account for nearly 90 percent of all malaria cases. The majority

of the remaining cases are clustered in India, Brazil, Afghanistan, Sri Lanka, Thailand,

Indonesia, Vietnam, Cambodia, and China. Malaria causes 1 to 1.5 million deaths each

year, and in Africa, it accounts for 25 percent of all deaths of children under the age of

five.

The commonly employed method for diagnosis of malaria involves the

microscopic examination of Romanowsky stained blood films. The peripheral blood smear

detects malarial parasites only when 40-60 parasites/microlitre of blood are present6. In

recent years numerous quick and new techniques for malaria diagnosis have been

developed, one such being the QBC (quantitative buffy coat) technique.QBC technique

detects malarial parasite even when they are only 1-2 parasites/microlitre of blood.

Many comparative studies of peripheral blood smear and QBC technique

are available in the literature. Conclusions of some of the studies are given below.

In a comparative study done by Manjunath P. Salmani et al, QBC method

was compared with thick and thin peripheral blood smears and malaria antigen test .

From this study they found that QBC method was superior to the smear for malarial

parasite detection, however species identification was not possible in majority of cases by

this technique. The study was concluded stating that though, the QBC method has its

advantages in terms of speed, sensitivity and ease, Leishman stained thin blood smear

appeared superior for species identification.3

Shujatullah F et al did a comparative study of different diagnostic

techniques in plasmodium falciparum cerebral malaria . In that they concluded that

sensitivity and specificity of ParaSight-F test were 96.6 and 94% while QBC was 97.8

and 100% respectively. ParaSight-F test and QBC were found to be novel methods for the

diagnosis of cerebral malaria especially in the cases where diagnosis can not be made by

conventional blood smear examination due to low parasitaemia. These rapid diagnostic

methods help in early therapeutic intervention.4

In a Comparative study of peripheral blood smear , quantitative buffy coat

and modified centrifuged blood smear ,Bhandari PL et al concluded that peripheral blood

smear had 86.79% sensitivity and was absolutely specific .QBC was 96.22% sensitive and

93.61% specific.5

In a comparitive study of MJW pinto et al QBC was compared with thick and thin

peripheral blood smears in 2274 samples. Malaria was diagnosed in 239(10.5%) Patients

by Leishman’s staining technique and QBC method . The QBC method allowed detection of

an additional 89 (3.9%) cases. Thus the prevalance rate of malaria during the study was

14.4%. In 1946, who were negative by the QBC technique , the Leishman’s stained smear

did not provide any help in malaria diagnosis. Analysis of the relative quantity of parasites

in the speciemens , in the QBC method, revealed that 80 out of 89 QBC positive but smear

negative cases had a very low parasite number (less than 10 parasites per QBC

field).Although the QBC method was superior to the smear for malarial parasite

detection,species identification was not possible in 26 (7.9%) cases by this technique . In

95.7% QBC positive cases ,they found that the buffy coat in the QBC tube appeared

pigmented (grey to black). The color of the buffy coat was therefore considered as a

predictor of positivity and could be taken as an indicator for a careful and more prolonged

search for the parasites. They suggested that the QBC technique has its advantages in

terms of speed sensitivity and ease. However , the Romanowsky stains still appear

superior for species identification.6

6.3 OBJECTIVE OF THE STUDY

To compare the efficiency of QBC and Leishman’s smear techniques (thick and

thin) in detection and identification of malarial parasite in suspected patients referred in a

tertiary health care hospital.

7. MATERIALS AND METHODS

7.1 SOURCE OF DATA

Two hundred malaria suspected patients referred for QBC in Father Muller Medical

College Hospital, Mangalore.

Inclusion criteria

All malaria suspected patients referred for haematological investigation in Father

Muller Medical College Hospital, Mangalore.

Exclusion criteria

Patients referred for other investigative procedures.

7.2 METHOD OF COLLECTION DATA

The blood sample of the malaria suspected patients will be

collected and assessed by using the technique of QBC. The same sample will be

assessed by using Leishman’s smear techniques (thin and thick). The results of the

sample analysis by using the three different techniques, will be documented and

assessed statistically. The statistical data will be interpreted to compare the efficiency of

QBC and Leishman’s smear techniques (thick and thin) in detection and identification of

malarial parasite.

7.3 Does the study require any investigations or interventions to be conducted on

patients or other human or animal?

No.

8. LIST OF REFERENCES

1) Mendiratta DK ,Dhutuda K. Narang R, Narang P. Evaluation of different for

diagnosis of P.falciparum malaria. Indian J Med Microbiol 2006;24(1):49-51.

2) Malik S , Khan S , Das A , Samantaray J.C , Plasmodium lactate dehydrogenase

assay to detect malarial parasites. Natl Med J India 2004;17:237-9.

3) Salmani MP, Mindolli PB , Peerapur BV.Comparitive study of peripheral blood

smear, QBC and antigen detection in malaria diagnosis . JCDR 2011;5(5); 967-969

4) Shujatullah F, Malik A , Khan HM & Malik A. Comparison of different

diagnostic techniques in plasmodium falciparum cerebral malaria.J Vect Borne Dis

2006;43;186-190

5) Bhandari PL , Raghuveer CV, Rajeev A ,Bhandari PD. Comparitive study of

peripheral blood smear, quantitative buffy coat and modified centrifuged blood

smear in malaria diagnosis.Indian J Pathol Microbiol 2008;51;108-112

6) Pinto MJW , Rodrigues SR , Desouza , Verenkar MP. Usefulness of quantitative

buffy coat blood parasite. Detecton system in diagnosis of malaria. Indian J med

microbiol 2001;19(4):219-221

9. SIGNATURE OF THE

CANDIDATE:

10. REMARK OF THE GUIDE:

11. NAME AND DESIGNATION OF

(in block letters)

11.1 GUIDE DR. HILDA FERNANDES.

PROFESSOR AND HEAD OF THE DEPARTMENT

DEPARTMENT OF PATHOLOGY

FATHER MULLER MEDICAL

COLLEGE, KANKANADY

MANGALORE – 575002

11.2 SIGNATURE

11.3 HEAD OF THE

DEPARTMENT

DR. HILDA FERNANDES.

PROFESSOR AND HEAD OF THE DEPARTMENT

DEPARTMENT OF PATHOLOGY

FATHER MULLER MEDICAL

COLLEGE, KANKANADY

MANGALORE – 575002

11.4 SIGNATURE

12. 12.1 REMARKS OF THE

CHAIRMAN AND DEAN

12.2 SIGNATURE