Download Hematological changes like anaemia are the most common

RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES
KARNATAKA, BANGALORE
ANNEXURE – II
PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION
JEETENDRA SHARMA
I YEAR M. Sc. MLT
DR. M. V. SHETTY COLLEGE OF MLT
VIDYANAGAR
MANGALORE – 575 013.
1.
Name of the candidate and address
(in block letters)
2.
Name of the Institution
DR. M. V. SHETTY COLLEGE OF MLT
VIDYANAGAR
MANGALORE – 575 013.
3.
Course of Study and Subject
M. Sc. MLT
HAEMATOLOGY AND BLOOD
TRANSFUSION
4.
Date of Admission to the Course
25-11-2009
5.
Title of the study
“COMPARISON STUDY OF THICK PERIPHERAL BLOOD SMEAR,
QUANTITATIVE BUFFY COAT AND MODIFIED CENTRIFUGED
BLOOD SMEAR IN MALARIA DIAGNOSIS”
1
INTRODUCTION
Malaria, sometimes called the “King of Diseases”, is caused by protozoan parasites
of the genus Plasmodium.1 The Plasmodium species are P. vivax, P. falciparum, P. ovale
and P. malariae. Among them P. vivax and P. falciparum are the most common infectious
agents in India. The most serious and sometimes fatal type of malaria is caused by
Plasmodium falciparum.2
Malaria can be transmitted mainly through mosquito bites, and by blood transfusion,
through needles and syringes in narcotic addicts.3
The malaria parasites start to multiply within red blood cells, causing symptoms that
include fever and headache. In severe cases, the disease worsens, leading to coma and death.
Hematological changes like anaemia are the most common complication
encountered in malaria and play a major role in the fatality.4
The two factors which play a crucial role in the development of anaemia are:
1.
Increased rate of red cell loss due to haemolysis.
2.
Defective production of red cells in bone marrow.
The present study is done to compare the use of thick peripheral blood smear,
Quantitative Buffy Coat (QBC) technique and Modified Centrifuged Blood Smear in
malaria diagnosis.
6.
Brief resume of the intended work:6.1 The need for the study
“Prevention is better than cure”.
Malaria is the most important infectious disease in tropical and subtropical regions,
and continues to be a major global health problem, with 300 to 500 million cases and among
them, 2 to 3 million deaths per year .5
2
According to WHO, half of the world’s population is at risk of malaria, and an
estimated 243 million cases led to nearly 8,63,000 deaths in 2008.6
Malarial parasites had their origin in Africa along with the mankind. However, about
90% of all malaria deaths in the world today occur in the sub-Saharan Africa.7
Malaria is one of the major public health problem in the India and according to
National Vector Borne Disease Control Programme (NVBDCP) around 1.5 million
confirmed cases are reported annually, of which 40–50% are due to Plasmodium
falciparum.8
In India, the North-eastern states are most affected, of which Assam tops the list in
terms of incidence and mortality. Malaria is also a problem in the states of
Andhra
Pradesh, Karnataka, Madhya Pradesh, Gujarat, Haryana, Punjab, and Orissa. The largest
focus of P. malariae in India is reported to be in Tumkur and Hassan districts of Karnataka. 9
In the year 2006, a total of 62864 cases of malaria were reported from Karnataka
state and Mangalore accounted for 15664 (24%) of these cases. Of the 16446 cases of P.
falciparum malaria reported from Karnataka in the same year, 4903 (29%) cases were from
Mangalore. And among 29 malaria related deaths from Karnataka, 11 were from
Mangalore.10
Malaria affects almost all the organs in the body. But one of the chief components
affected is blood. This study is done to compare the efficiency of different method in
diagnosis of malaria. Since the data available on the use of Modified Centrifuged Blood
Smear (MCBS) is sparse, this method is compared with other well established method like
Quantitative Buffy Coat (QBC) technique in the diagnosis of malaria.
The above data shows that malaria is a major problem. Hence it is necessary to find
a reliable and valuable method for diagnosis that can be applicable for a large population.
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6.2
REVIEW OF LITERATURE
“Malaria particularly falciparum malaria is a major public health problem in India.
Its correct and early diagnosis is very important for prompt treatment as a preventive and
control measure. Microscopy is the traditional method for laboratory diagnosis of malaria,
which is used widely. However, it is time consuming, needs expertism and the detection
limit is 10-20 parasites/ml blood in thick film and 100 parasites/ml bloods in thin film.
Quantitative Buffy Coat (QBC) technique is highly sensitive method but expensive
equipment is needed for this test. The serological methods involving antibody detection
give information regarding exposure to malaria but do not differentiate between present and
past infections. 11
In the study conducted by P.L. Bhandari on patients from Wenlock Govt. Hospital, it
was found that modified centrifuged blood smear (MCBS) was successful in detecting
nearly 100% of malaria cases and Quantitative Buffy Coat (QBC) was positive in only
96.22% .12
In the study conducted in JN Medical collage hospital, Aligarh, Quantitative Buffy
Coat (QBC) technique was able to detect only 46 out of 50 of cerebral malaria.13
A study on comparison of different diagnosis technique in malaria was carried out in
Kempegowda Institute of Medical Science and Research Hospital Bangalore. 102 suspected
patients of malaria were chosen for study. Among them 62 cases were positive by peripheral
blood smear (PBS) and 76 cases were positive by Quantitative Buffy Coat (QBC) method.14
In the study conducted by Jain M and Kaur M, 2005 in diagnosis of malaria was
carried out. A total 200 blood samples are chosen for study. Among them 70(35%) samples
were positive by Quantitative Buffy Coat (QBC) technique, 62(31%) sample by thick
smear, 50(25%) samples by thin smear and only 34(70%) samples were positive by
conventional buffy coat technique.15
In the study by Parija SC the efficacy of various methods like i.e.; thick and thin
smear and Quantitative Buffy Coat (QBC) in diagnosis of malaria was carried out. A total of
411 samples were collected from patients presenting with classic symptoms of malaria. For
traditional microscopy; thick and thin smear were prepared and stained with Leishman’s
4
stain, taking thick smears as gold standard, thin smear had a sensitivity and specificity of
54.8% and 100%, respectively. Quantitative Buffy Coat (QBC) show 78%. Leishman’s
thick smear, although cost effective, is difficult to interpret for inexperienced; so if facilities
are available, Quantitative Buffy Coat (QBC) should be used for routine diagnosis.16
In the study by Krishna BV and Deshpande AR, 2003 on the comparison between
conventional and Quantitative Buffy Coat (QBC) method for diagnosis of malaria. 1435
blood samples were chosen for study. it was found that 57(3.97%) samples were positive by
Quantitative Buffy Coat (QBC) technique while only 34(3.07%) samples were positive by
blood film.17
A study on comparison of QBC with both the thin and thick film in the diagnosis of
malaria among three groups of hospitalized patients. The Quantitative Buffy Coat (QBC)
technique test showed more sensitivity than the two conventional methods in the three
groups as follows: in group (1) the Quantitative Buffy Coat (QBC) technique was positive
in 14.2% compared to 9% only in either thin or thick blood films. In group (2) the
positively was 95.1% compared to 79.3% & 76.8% in the thin and thick blood films
respectively. In group (3) the sensitivity was 22.9% compared to only 5.7% in both of the
thin and thick blood films.18
Therefore, the study concluded that Modified Centrifuged Blood Smear (MCBS) and
Quantitative Buffy Coat (QBC) was getting more sensitive diagnosis for malaria compared
to peripheral blood smear (PBS).
6.3
AIMS/OBJECTIVE
Aims and objectives of this study are
1.
To screen the peripheral blood smear of patients with clinical diagnosis of malaria.
2.
To screen the same blood sample with rapid diagnostic test Quantitative Buffy Coat
(QBC) technique & Modified Centrifuged Blood Smear (MCBS).
3.
To compare the diagnostic and prognostic utility of rapid test with conventional
thick and thin films.
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7.
MATERIAL AND METHOD
7.1
SOURCES OF DATA:
Blood sample will be collected from 100 patient attending malaria clinics at
Government Wenlock Hospital and Dr. M.V Shetty College Hospital, Mangalore, during the
period from 2010 to 2011.
7.2
METHOD OF COLLECTION OF DATA:
Sample and sampling technique
Type of study: Purposive data collection.
Sample and sampling techniques: 100 malaria patient cases will be studied using purposive
sampling techniques
1.
Thick blood smear by Leishman’s stain or Giemsa stain examination to detect the
presence and type of malaria.
2.
By Quantitative Buffy Coat (QBC) technique and Modified Centrifuged Blood
Smear (MCBS) rapid diagnosis test:
In order to perform QBC technique 50 micro liter of whole blood is taken into a
capillary tube coated with acridine orange and fitted with a cap. A plastic float is inserted
inside the tube and centrifuged at 1200 rpm for 5 min. After centrifuge the tube is mounted
on a small plastic holder and examined through an ordinary fluorescence light microscope.
For Modified Centrifuged Blood Smear (MCBS) centrifuged capillary tube is cut
and simultaneously blood smear is made.
7.2.1 Sample size
The sample size consists of 100 malaria patients at Government Wenlock Hospital
and Dr. M.V Shetty college Hospital, Mangalore.
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7.2.2 Inclusion criteria for sampling

All adult patients with a positive blood smear for P. falciparum Malaria or
Quantitative Buffy Coat (QBC).

Those who are ready to be a subject matter.
7.2.3
Exclusion criteria for sampling

Paediatric patients

Patients who are peripheral smear negative but treated with anti malarial drugs (so
called clinical malaria).

Other malarias (Plasmodium ovale, Plasmodium malariae).
7.2.4 Instruments intended to be used

Demographic data.

Laboratory procedures.
7.2.5 Plan for data analysis

Data will be analyzed using purposive sampling technique by blood smear,
Quantitative Buffy Coat (QBC) technique & Modified Centrifuged Blood Smear
(MCBS) technique.

Findings will be presented using tables and figures.
7.3
Does the study require any investigations or interventions to be conducted on
patients, or other animals? If so please describe briefly.
Yes, the study requires drawing blood sample from suspected malaria patients.
7.4
Has ethical clearance been obtained from your institution in case of 7.3?
Permission to conduct the study is obtained from the institution. Permission will be
obtained from the concerned authority at the time of data collection.
7
8.
List of References
1.
Curing malaria together. Korean J Parasitol 2009 Jun;47(2):93-102.
2.
Reddy RK. Text book of microbiology and parasitology. 2nd ed; 2005.
3.
Deodhare SG. Infections by animal parasites. In: General Pathology and Pathology
of Systems. Ed: Deodhare SG, 6th ed. Vol. 2; 2002.
4.
Khaled T, El-Dein SZ, Majid I, et al. Haematological change in malaria: Relation to
plasmodium species. Kuwait Medical Journal 2007;39(30):262-7.
5.
Aslan G, Ulukanligil M, Seyrek A, Erel O. Diagnostic Performance Characteristics
of Rapid Dipstick Test for Plasmodium vivax Malaria. [online]. Available:
URL:memorias.ioc.fiocruz.br/965/4147.pdf
6.
The World malaria report 2009. [online]. Available from:
URL:www.who.int/malaria/world_malaria...2009/en/index.html
7.
Snow RW, Guerra CA, Noor AM, Myint HY, Hay SI. The global distribution of
clinical episodes of Plasmodium falciparum malaria. Nature 2005;434(7030):214-7.
8.
Diagnosis and Treatment of Malaria in India. [online]. Available from:
URL:www.mrcindia.org/Guidelines_for_Diagnosis___Treatment.pdf
9.
Park K. Malaria. In: Park’s Textbook of Preventive and Social Medicine. Ed: Park
K. 19th ed. Jabalpur: Banarsidas Bhanot; 2007.
10.
Malaria in Mangalore. [online]. Available from: URL:
www.malariasite.com/malaria/MalariaInMangalore.htm
11.
Thakor HG. Laboratory diagnosis of malaria. J Indian Med Assoc 2000;98(10):6237.
12.
Bhandari PL. Comparative study of peripheral blood smear, Quantitative buffy coat
and modified centrifuged blood smear in malaria diagnosis. [online]. Available
from: URL:http://www.ijpmonline.org/pubmed
8
13.
Shujatullaha F. Paediatric and medicine wards of J. N. Medical College Hospital,
Aligarh, India. [online]. Available from:
URL:www.mrcindia.org/journal/issues/434186.pdf
14.
Chandrakanth GC. Comparison of conventional blood film method with QBC and
parasite lactate dehydrogenase, Histidine Rich protin-2 the rapid diagnosis of
malaria. Unpublished Doctoral dissertation submitted to Rajiv Gandhi University of
Health Sciences, Bangalore; 2006.
15.
Jain M, Kaur M. Comparative study of microscopic detection methods and
haematological changes in malaria. Indian J Pathol Microbiol 2000;48(4):464-7.
16.
Parija SC, Dhodapkar R, Elangovan S, Chaya DR. A comparative study of blood
Smear, QBC and antigen detection for diagnosis of malaria. Indian J Pathol
Microbiol 2009 Apr-Jun;52(2):200-2.
17.
Krishna BV, Deshpande AR. Comparison between conventional and QBC methods
for diagnosis of malaria. Indian J Pathol Microbiol 2003;46(3):517-20.
18.
el Serougi AO, Amin AM. The quantitative buffy coat capillary tubes versus thin
and thick blood films in the diagnosis of malaria in Saudi Arabia. J Egypt Soc
Parasitol 1998;28(1):17-22.
9
9.
Signature of the candidate
10.
Remarks of the guide
11.
Name and designation of (in block letters)
11.2 Guide
DR. SUCHITHRA . K . RAO
PROFESSOR
MBBS , MD (PATHOLOGY)
DR . M . V . SHETTY COLLEGE OF MLT
VIDYANAGAR , MANGALORE – 575013
11.2 Signature
11.3 Co-guide (if any)
11.4 Signature
12
12.1 Head of the department
DR. SUCHITHRA . K . RAO
PROFESSOR
MBBS , MD (PATHOLOGY)
DR . M . V . SHETTY COLLEGE OF MLT
VIDYANAGAR , MANGALORE – 575013
12.2 Signature
13.
13.1
Remarks of the Chairman and Principal
13.2
Signature
10