Frequency of Hepatitis C Virus in post-surgical patients

Submitted By: Supervisor:

Ahmad Ali Zahid Dr Imtiaz Mehmood Tahir
Roll No 21213 BSc (Hons) MLT, M.Phil.
B.s (Hons) MLT PhD Bio Chemistry

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 Introduction

INTRODUCTION

Chronic HCV is the main reason of liver disease and cirrhosis, and the main indicator of
liver transplant in the Europe. Round about 171 million individuals were affected with
hepatitis C virus in the whole world, consisting of 4% of the global population. Hepatitis
C virus was the main chronic blood borne infection in the United States. and was take
part in 41% of chronic liver abnormality. It was detected from the serum of a individual
with non-A, B hepatitis initially in 1989 by Cholet al. shortly after cloning of HCV, this
newly identified virus was detected to be the reason of round about 91% of non-A-B
hepatitis in the United States. This is an RNA virus that have a relation with the family
Flaviviridae (Stephen. L Chen, 2006).

1.1 Susceptibility of infection

Susceptibility is common. Human and chimpanzees are the only one known susceptible
that infect, with species developing same disease.

1.2 Causes of disease

HCV is mainly due to viral infection with the HCV, an enveloped, single stranded,
positive sense RNA virus. The virus infects hepatocytes and can cause severe
inflammation of the liver with long term complications. The progress of disease is
generally imsidous, with anorexia, abdominal pain, nausea and vomiting. Out of these
exposed persons to HCV, round about 41% recover completely, whether the remaining,
may or may not have symptoms, can become chronic liver carriers. Of these, 21% can
develop liver cirrhosis. Those who have liver cirrhosis, up to 21% to develop liver
carcinoma.

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 Introduction

1.3 Vaccine For HCV

Currently there is no Vaccine for HCV because virus come in many forms.

1.4 Classification

The Hepatitis C infection has a place with the different types of hepatitis C virus an
individual from the family of Flaviviridae.

1.5 STRUCTURE

The HCV molecule comprises of a center of hereditary material RNA.

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 Introduction

Genome

HCV has a +vee sense single stranded RNA genome. The genome comprises of a solitary
Open casing is 9600 Nucleotide long Bases.

Medical Importance

Genotype is medically imperative in deciding the reaction to interferon-based prognosis

and the given period of time of such treatment.

1.6 Epidemiology

Hepatitis C infection is prevalently a blood-born infection, with sexual or vertical

transmission. As a result of this method of spread the key gatherings at danger are
infusing medication clients, individuals that have Transfused Blood, beneficiaries of
blood items and at times patients on hemodialysis. Regular settings for HCV inoculation
is

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 Introduction

additionally, Intra Healing facility transmissions, whenever practices of cleanliness and

cleansing never accurately follow in the center.

1.7 Stability in the environment

In the same path as other infections, the HCV infection is continued to inactivate outside
of the body of host. The vicinity of warmth could extraordinarily decrease the lifespan of
the infection outside the body. This infection could stay with out the resistance outside a
host for round about the sixteen days at 25°C and two days at 36°C while it could stay
dynamic for six weeks at temperatures not as much as or equivalent to 4°C. At the point
when warmed to temperatures of 60°C and 65°C, be that as it may the hepatitis C
infection could be inactivated in eight and four mines of individual.

Transmission.

HCV is blood borne viral infection. It is mostly transmitting through:

Injection of drug by using the share of injecting equipment.

In the clinical care setting because of re-use or poor sterilization of clinical equipment
mainly syringes and needles.

The transfusion of blood and blood products without screening.

Hepatitis C virus could be transmitted through sexual contact and could be pass from
the mother with infection to her baby.

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 Introduction

Symptoms

The Acute HCV symptoms

A small group of people that have HCV Infection will get sick with acute hepatitis C.
Acute HCV symptoms are loss of hunger, early vomiting, fatigue, temperature,
abdominal discomfort and jaundice. Acute HCV is generally a less illness from which
many of them recover within three to eight weeks.

Chronic HCV symptoms

Chronic HCV carriers are at most risk of long-term liver disease, which is due to
increasing damage of liver. At the initial stage symptoms are lower fatigue, a feeling
abnormality, and intolerance of alcohol. Severe conditions of long-term liver disease are
scarring of the liver, it may not become visible upto 21 to 50 years after getting infection
with the virus. It may can cause liver failure or liver carcinoma.

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 Introduction

1.14 Extra Hepatic diseases.

1: Cryoglobulinemia
2: Glomerulonephritis
3: Porphyria Cutanea tardy
4: Lichen Planus
5: Diabetic Nephropathy
6: B-Cell Non-Hodgkin Lymphoma
7: Autoimmune Thyroiditis
Pathogenesis

1; Innate Immune Response to HCV

2; Humeral Immune Response to HCV

Cellular Response to HCV

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 Introduction

Transmission of HCV in the Clinical-care setting

HCV-RNA can also be detected in ascitic fluid, semen in addition to blood detection and.
HCV urine positive patients with chronic liver disease. The Transmission of HCV
infected to clinical-care persons has commonly been documented following percutaneous
exposure of blood. Some cases are reported of transmission by mucous membrane
exposure, through a blood splashes to the conjunctiva.

Transmission from Mother to Infants

It is reported in several cases the transmission from mother to infants, Detection is
determined by HCV-RNA PCR in the child range from 0 to 7 percent. During C section
in operation theatre can transmit the HCV-RNA virus (M. MacDonald, 1996).

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 Literature Review

LITERATURE REVIEW

Case no. 1

Alexandra E. et al in 1991 give a study to identified the prevalence of anti HCV in

patients whose undergone liver-transplant. In this study 128 patients who were liver
transplantation. The 66 patients who have 6 months of follow up and post- transplant
samples were entered in the study for Anti HCV antibody detection. This study shows
Hepatitis Virus is a due to Chronic Liver Disease in patients who were undergone Liver
Transplantation. (Alexandra E. Read, et al., 1991)

Case no.2

Takuya Komura et al (2007). conducted a study on diabetic patients for the determination
of post-operative HCC recurrence with HCV infection. This study was the controversial.

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Literature Review

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 Literature Review

2.6 Objective:

The objective of study was to determine the frequency of Hepatitis C Virus in patients who
undergone any surgical procedure.

2.7 Rationale:

The rationale of my study is whether surgery can be the possible cause of hepatitis C

infection.

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 MATERIALS AND METHODS

3.1 Study Population

Patients were included in this study that had undergone surgery.

3.2 Clinical settings

This present study was transfer at the Department of Pathology, FMH college of
Medicine & Dentistry, Lahore and the samples were collected from Mayo Hospital
Lahore. This study was started with the supervision and permission of Mr. Abubakar
Imran, H.O.D MLT FMH Institute of Allied Health Sciences and Mr. Muhammad
Ibrahim Associate Professor of Statistics, FMH Institute of Allied Health Sciences.

3.3 Study Design

This study was an Observational Descriptive study. This study did not involve any kind
of treatment.

Sample size

Blood samples of about 105 patients were collected from Mayo Hospital
Lahore. (UMAIRA ALAUDDIN, 2007)

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 3.4 Inclusion criteria

Post-operative patients with HCV positive and HCV negative by PCR in both male and
female were included in this study.

3.5 Data Collection Instrument

Patient samples were selected from Surgical Ward, Mayo Hospital Lahore. The
questionnaire which was had all the possible queries regarding the study. Blood samples
were collected under aseptic conditions provided by spirit swab using 5 ml BD syringes.

3.6 Procedure

The samples were collected from Mayo Hospital Lahore, using aseptic technique. The
procedure was HCV PCR by Real Time PCR method.

3.9 Ethical issues

There were no ethical issues regarding this study.

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 3.9.1 Polymerase chain Reaction (PCR) steps

PCR is basic molecular technique which is used for amplification of the required gene in three
steps.

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3.9.2 Gel Electrophoresis steps

1. 1xTAE (Tris-acetate-EDTA) was prepared.

2. Agarose gel was prepared by melting agarose and 1xTAE in the microwave; time was
given to cool down.

3. When the gel was cooled, then it was placed in the electrophoresis
equipment and comb was removed.

4. PCR products were collected from the fridge.

5. Spots of loading dye were made on para film.

6. Sample was added to a spot, mixed it.

7. Previous step was repeated for each sample and for a –vee control.

8. PCR Ladder was loaded and mixed with loading dye in to well.

9. Electrophoresis apparatus was connected to a power supply.

10. Power was adjusted to run 80-120 V.

11. Waited 20-30 minutes.

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 12. Power was disconnected and collected the gel carefully by the liquid.

13. Gel was observed carefully.

3.12 Statistical Analysis:

The categorical values were expressed in the form of frequency. Tables and graphs were

used to display the data. Appropriate statistical tools were used to analyze the data.

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 Results

Results

A total number of 105 samples were analyzed by Polymerase Chain Reaction method.
Out of 105 patients among them 73 samples (69.5%) were males and 32 samples (30.5%)
were females.

Frequency Percent

Male 73 69.5
Female 32 30.5
Total 105 100.0

Table No.4.1: Gender distribution

80
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50
Frequency
40
30
20
10
0
Male Female

Graph no. 4.1: Distribution of gender

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 Results

Out of 105 samples 17 samples (16.2%) were positive and 88 samples (83.8%) were
negative by Polymerase Chain Reaction method.

Frequency Percent

Positive 17 16.2
Negative 88 83.8
Total 105 100.0

Table No.4.2: HCV by PCR

90
80
70
60
50 Frequency
40
30
20
10
0
Positive Negative

Graph No.4.2 HCV by PCR

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References

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 References

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Olsen, D. P. (2010). Increased risk of transmission of Hep C in open heart surgery

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