This document discusses a study that will examine the incidence of diabetes mellitus in patients with hepatitis B and C virus infections. The study will collect blood samples from 502 patients who test positive for HBV or HCV. PCR and HbA1c tests will be performed to confirm the viral infections and presence of diabetes. It is hypothesized that abnormal glucose tolerance should be seen in chronic liver disease, as the liver plays a role in glucose metabolism. The results of this study could help understand the relationship between hepatitis viruses and diabetes.

2. Muhammad
Salman Siddique
Al-1161
B.S.(Hons.) Medical Lab Technology Final Prof.
Fatima Memorial Institute of Allied Health Sciences
Shadman Lahore

3. Topic
Incidence of Diabetes Mellitus in patients
with Hepatitis B and C virus infection.

4. Introduction
Hepatitis C Virus (HCV) infection is one of the most
important cause of cirrhosis worldwide. (Simona, et al.,
1999) Although HCV targets at the liver, it has become
interestingly evident that HCV can induce diseases of
many organs. It is reported that 38% of patients with
HCV would manifest at least one extrahepatic manifestation
during the illness. (Eman, et al., 2011) There are also
some other evidences that both HBV and HCV have extra
hepatic sites of viral colonization resulting in extrahepatic
manifestations especially of kidneys and pancreas resulting
in proteinurea and diabetes. (Qureshi, et al., 2002)

5. Introduction (cont.)
Association of diabetes mellitus with chronic liver disease
has been recognized for over 2 decades and the following
hypothesis has been suggested:
“As liver is involved in glucose metabolism
therefore it is presumed that abnormal glucose tolerance
should be seen in chronic liver disease and this was
confirmed in many studies, whereas about 70% cirrhotics
were found to have abnormal glucose tolerance. (Qureshi,
et al., 2002)”

6. Introduction (cont.)
The highest Hepatitis C Virus (HCV) prevalence in the
world occurs in Egypt at an estimated 12% among the
general population and reaches 40% in persons 40 years
of age and above in rural areas. (Eman, et al., 2011) In
Paskistan the HBV carrier and HCV exposure rates are
high; being 8-10% for HBV and 4% for HCV. (Qureshi, et
al., 2002)

7. Objectives
• To observe the association of Diabetes Mellitus with
patients suffering from Hepatitis B and C virus infection.
• To magnify the relationship between Diabetes Mellitus
and Hepatitis B and C virus infection to spread the
awareness of their association.

8. Rationale
The purpose of this study is to conclude the relationship
of Diabetes Mellitus with Hepatitis B and C. It will also
help magnify the problem and co-existence of Diabetes
Mellitus and Hepatitis so that the treatment of Hepatitis B
and C can be optimized.

9. Materials and Methods
Research work location:
This present study will be conducted at the Department
of Pathology, FMH College of Medicine & Dentistry,
Shadman Lahore.
Sample Size:
Approximately 502 mixed samples of HCV and HBV
positive patients will be collected. (Akbar, Siddique &
Ahmad, 2002)

10. Materials and Methods (cont.)
Inclusion criteria:
Patients positive of HBV or HCV infection will be
included.
Exclusion criteria:
Patients negative of HBV or HCV infection will be
excluded.
Study Design:
Observational study.
Duration:
Six months.

11. Materials and Methods (cont.)
Data Collection Instrument
Questionnaire will be filled according to the collected
information of inclusive individuals.
Data Analysis:
Data will be entered and analyzed using SPSS 17.0.
(SPSS Inc. Chicago, USA). Appropriate statistical tools
will be applied to analyze the data.
Ethical Issues:
There will be no ethical issue observed during this
study.

12. Methodology
Sampling: Routine blood sampling technique will be used to
collect 3ml blood sample in clotted vial for PCR and 3ml
in EDTA anticoagulant vial for HbA1c from patients that
will be included in this study. The samples will be stored
at 2-8˚C in a refrigerator.
PCR: Then PCR will be done to confirm the viral
infection. PCR depends on three steps denaturation,
annealing and elongation. PCR is done by repeating
several thermal cycles in a controlled manner. In
denaturation step the DNA is heated usually to 95oC to
render it single stranded.

13. Methodology (cont.)
Then in annealing step the two primers bind the
appropriate complimentary strand. And in elongation step
DNA polymerase extends the primer by its polymerase
activity. This is done at a temperature optimal for the
particular polymerase that is used. Currently the most
popular enzyme for this step is Taq Polymerase, the DNA
polymerase from the thermophilic (heat loving) bacteria
Thermus aquaticus . The elongation takes place at 72oC.
These steps are repeated from 28-35 times. In PCR the
desired genetic material is enhanced and then its load is
measured.

14. Methodology (cont.)
HbA1c: HbA1c test will be performed to confirm the
Diabetes Mellitus according to the WHO’s Diagnostic criteria
for Diabetes Mellitus, in those patients that are confirmed
of having Hepatitis infection in PCR. Latex enhanced
immunoassay method of HbA1c will be used. It is based
on interaction between antigen molecules (HbA1c) and
HbA1c specific antibodies coated on latex beads. This
cross link reaction results in changes in the solution
turbidity which is proportional to the amount of the antigen
in the sample.

15. References
• Eman, L. E., Gamal, F. M., Mai, A. E.-D., Fatma, A.
M., Gamal, G. E., & Mohamed, A.-h. (2011). Association
of HCV with diabetes mellitus: an egyptian case control
study. Virology, 367(8).
• Qureshi, H., Ahsan, T., Mujeeb, S., Jawad, F., Mehdi, I.,
Ahmad, W., & Alam, S. (2002). Diabetes mellitus is
equally frequent in chronic HCV and HBV infection. J
Pak Med Association, 52(7), 280-283.

16. References (cont.)
• Simona, C., Kevin, T., Luigi, P., Colin, C., Ugo, P.,
Juraj, P., Graeme, J. A. (1999). Further Evidence for an
Association between Non–Insulin-Dependent Diabetes
Mellitus and Chronic Hepatitis C virus infection.
Hepatology, 30(4), 1059-1063.
• Akbar, D., Siddique, A., & Ahmad, M. (2002). Diabetes
mellitus is equally frequent in chronic HCV and HBV
infection. Med Princ Pract, 11(2), 82-85.

17. Incidence of Diabetes Mellitus in patients with Hepatitis B
and C virus infection.
Serial no. ________
Name: ______________________________
Age: _______
Gender: ____________
Occupation: ____________________
PCR test report:
Detected Not Detected
HBV
HCV
HbA1c test report:
Date: __/__/201_ __________
Signature
Control Reference
Value
Patient
Value
Ideal/Excellent 4.0 – 6.0 %
Desirable 6.1 – 7.0 %
Suboptimal 7.1 – 8.0 %
Very poor > 8.1 %