NIPER-Hajipur

METHODS

MATERIAL AND

4. MATERIAL AND METHODS

Study site: The study was conducted in out-patient department of RMRIMS (Rajendra
Memorial Research Institute of Medical Science), ICMR (Indian Council of Medical
Research), Agamkuan, Patna, India.
Study design: The study was prospective cross-sectional study.
Study duration: From August 2014 to May 2015.
Ethical approval: The clinical protocol entitled A prospective cross-sectional study to
assess prevalence of cardiovascular risk factors in Hypertensive and Diabetic patients
was approved by Ethical Review Board, RMRIMS, PATNA.
Study Criteria:
Inclusion Criteria:
More than 20 years age group.
Patients diagnosed with diabetes or hypertension of both sex.
Patients who were not willing to give consent for study.
Exclusion criteria:
Age less than 20 years.
Previously diagnosed HIV and Hepatitis patients.
Patients who were not willing to give their consent.
Plan of work:
We have planned to study CV risk factors among 200 diabetic and hypertensive patients.
Data was collected using a risk factor assessment form (questionnaires).The risk factor
assessment form had four parts. The first section was related to socio-demographic
characteristics of the patient. The second section was about habits including cigarette
smoking, alcohol drinking, tobacco chewing and information related to family history of

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NIPER-Hajipur
METHODS

MATERIAL AND

diabetes, hypertension or cardiovascular disease. The third part was for clinical and
laboratory results. In the final part anthropometric measurement like (height, weight,
waist circumference, hip circumference, BMI, blood pressure was recorded. Weight of
each patient was measured in( kg) using WHO weighing scale. Height was measured in
cm in erect position with shoes removed. Waist circumference and hip circumference was
also measured using a tape. Blood pressure was measured using mercury
sphygmomanometer in sitting position. Hypertension was defined as blood pressure
140/90 mmhg. BMI was calculated using standard formula.
BMI was calculated as weight (kg) divided by the square of the height (m)
BMI (Kg/m) = weight in kg/height in meter
For laboratory examination purpose 5 ml of blood was taken in the laboratory of
RMRIMS after an overnight fasting of 10-12 hours.The patient attending the OPD were
advised to come after an overnight fasting for the next day for the determination of
Fasting blood sugar,serum choletsterol,C-rective protein,homocysteine ,antiphospholipid
level.FBS, Choletsteol, c-reactive protein were determined using Merck Microlab 300
autoanalyser and with appropriate reagents.
4.1 Determination of C-reactive protein level:
CliniQuant-TIA CRP Kit was used for the quantitative determination of C - reactive
protein in human serum by turbidimetric immunoassay. The reference value of CRP was
0-1 mg/dl with CliniQuant-TIA CRP Kit.
4.2 Determination of homocysteine level:
The Diazyme Homocysteine 2 Reagent Enzymatic assay Kit was used for quantitative
determination of homocysteine in serum.
4.3 Determination of antiphospholipid antibodies:
Antiphospholipid antibodies was determined using Antiphospholipid screen IgG/IgM
ELISA kit. The antiphospholipid screen IgG/IgM assay was used to screen the presence
of IgG and IgM class antiphospholipid antibodies in serum.

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NIPER-Hajipur
METHODS

MATERIAL AND

The procedure was followed as mentioned in ELISA Kit and reading was taken on Bio
Rad ELISA reader. Standard curve of IgG/IgM antibodies was plotted using MS EXCEL.
The graph was plotted between average optical density versus concentration. Then
Concentration of unknown was calculated from calibration curve by interpolation.
Normal antiphospholipid antibodies ranges:
IgG (GPLU/ml)

IgM (MPLU/ml)

Normal:

<10

<10

Elevated :

>10

>10

Statistical Analysis: All the data was analyzed using SPSS

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