Comparative analysis of HCV RNA, Anti-HCV and liver Transaminase levels:

predominance of HCV genotype

Fahmida NasrinID No: 1030883070MS in BiotechnologyDept. of Biochemistry & MicrobiologyNorth South University

Background

Hepatitis C Virus

At least 50% of patients develop chronic liver disease. Cirrhosis develops in about 20-30% within 5-30years. 15% develops hepatocellular carcinoma. Assess baseline viral load prior to initiation therapy Determine therapeutic success during and after therapy. Assessment of prognosis and disease progression.

Rationale of the study

Replication of HCV

Route of transmission Percentage

Injection Drug use 60%

Sexual 15%

Transfussion 10%

Occupation 4%

Other 1%

Unknown 10% 0%

10%

20%

30%

40%

50%

60%

70%

60%

15%10%

4% 1%

10%

Percentage

Transmission

Serologic Pattern of Acute HCV Infection with Recovery

Serologic Pattern of Acute HCV Infection with Progression to Chronic Infection

Epidemiology of Hepatitis C

Aims of the study This study was carried out : 1. to correlate serological marker of HCV and liver function

tests as an alternative to HCV RNA estimation. 2. to evaluate the state of infection among Anti-HCV positive

patients. 3. to decide the indication of therapy 4. to find out discordance of seromarkers with HCV RNA.

Materials and Method

Place of the study The study was conducted in- DNA Solution Ltd. Bangladesh Institute of Research and Rehabilitation in Diabetes, Endocrine and Metabolic Disorders (BIRDEM), Dhaka.

Study period This study was done during the period of April 2011 to November 2011.

Study design Cross sectional study.

Subjects

-Inclusion criteria Subjects with age ranging from 15-80 years. 120 patients diagnosed as a case.

-Exclusion criteria Patients with serious co-morbid disease (severe infection, stroke, myocardial onfarction, major surgery, mal-absorption etc).

Laboratory Analyses

Anti-HCV: Enzyme Linked Immunosorbent Assay (ELISA), Fortress Diagnostic-USA.

Serum ALT, AST: enzymatic assay, Randox-Germany. HCV RNA: Real-time Reverse Transcription PCR,

Roboscreen-Germany. HCV genotyping: Real-time Reverse Transcription PCR,

Sacace-Italy.Statistical Analysis All data were analyzed by using Statistical Package for

Social Science (SPSS) for Windows version 12.0. p value <0.05 was taken as level of significance.

Results

Age & sex distribution of subjects

Male Female0.00%

10.00%

20.00%

30.00%

40.00%

50.00%

60.00%

70.00%

80.00%

% of subjects

Sex No. of subjects (%)

Male86 (71.66)

Female34 (28.33)

Age (years) No. of subjects (%)<20 8 (6.66)

21-40 68 (56.66)41-60 42 (35)61-75 2 (1.66)

<20 21-40 41-60 61-750.00%

10.00%

20.00%

30.00%

40.00%

50.00%

60.00%

% of subjects

Age (years)

Prevalence of HCV infection in different age groups in male

0-10 11 to 20 21 to 30 31 to 40 41 to 60 61 to 7505

10152025303540

No. of male subjects

Age group No. of male subjects0-10 0

11-20 621-30 3531-40 1741-60 2661-75 2

Prevalence of HCV infection in different age groups in female

Age group No. of female subjects0-10 0

11-20 221-30 1831-40 1041-60 361-75 1

0-10 11 to 20 21 to 30 31 to 40 41 to 60 61 to 7502468

101214161820

No. of female subjects

0-10 11 to 20 21 to 30 31 to 40 41 to 60 61 to 7505

10152025303540

No. of male subjectsNo. of female subjects

Age group No. of male subjects

No. of female subjects

0 -10 0 011-20 6 221-30 35 1831- 40 17 1041- 60 26 361-75 2 1

Prevalence of HCV infection in different age groups in male & female

Correlation between Anti-HCV & HCV RNA

p=0.692

Positive HCV RNA Negative HCV RNA0.00

10.00

20.00

30.00

40.00

50.00

60.00

70.00

80.00

90.00

100.00

68.91

31.09

100.00

0.00

Positive Anti-HCV Negative Anti-HCV

 Positive Anti-HCV n=119

(%)

Negative Anti-HCV, n=01 (%)

Positive HCV RNA 82 (68.91) 1 (100.0)

Negative HCV RNA 37 (31.09) 0 (0.0)

Correlation between Anti-HCV and ALT

p=0.492

Raised ALT Normal ALT0.00

10.00

20.00

30.00

40.00

50.00

60.00

70.00

80.00

90.00

100.00

48.74 51.26

100.00

0.00

Positive Anti-HCV Negative Anti-HCV

 Positive

Anti-HCV, n=119 (%)

Negative Anti-HCV, n=01 (%)

Raised ALT 58 (48.47) 1 (100.0)

Normal ALT 61 (51.26) 0 (0.0)

Correlation between Anti-HCV and AST

p=0.433

Raised AST Normal AST0.00

10.00

20.00

30.00

40.00

50.00

60.00

70.00

80.00

90.00

100.00

42.86

57.14

100.00

0.00

Positive Anti-HCV Negative Anti-HCV

 Positive

Anti-HCV, n=119 (%)

Negative Anti-HCV, n=01 (%)

Raised AST 51 (42.86) 1 (100.0)

Normal AST 68 (57.14) 0 (0.0)

Correlation between HCV RNA and ALT  

p=0.549

Raised ALT Normal ALT40.00

42.00

44.00

46.00

48.00

50.00

52.00

54.00

56.00

49.40

50.60

45.95

54.05

Positive HCV RNA Negative HCV RNA

 Positive

HCV RNA, n=83 (%)

Negative HCV RNA, n=37 (%)

Raised ALT 41 (49.40) 17 (45.95)

Normal ALT 42 (50.60) 20 (54.05)

Correlation between HCV RNA and AST

p=0.425

Raised AST Normal AST0.00

10.00

20.00

30.00

40.00

50.00

60.00

42.17

57.83

45.96

54.05

Positive HCV RNA Negative HCV RNA

 Positive

HCV RNA, n=83 (%)

Negative HCV RNA, n=37 (%)

Raised AST 35 (42.17) 17 (45.96)

Normal AST 48 (57.83) 20 (54.05)

Frequencies of liver transaminases with HCV RNA & Anti-HCV

HCV RNA

  

Positive (n=83) Negative (n=37)

ALT+AST ALT AST ALT+AST ALT AST

Raised 33 (39.76%) 8 (9.64%) 2

(2.41%)15

(40.54%)3

(8.11%) 2 (5.41%)

Normal 40 (48.19%) 17 (45.95%)

Anti-HCV

  

Positive (n=119) Negative (n=01)

ALT+AST ALT AST ALT+AST ALT AST

Raised 47 (39.50%) 11(9.24%) 4 (3.36%) 1 (100.0%) 0 (0.0%) 0 (0.0%)

Normal 57 (47.90%) 0 (0.0%)

Viral Load 0 1-10^3 10^4 10^5 10^6 10^7 10^8

Raised ALT 19 (51.35) 5 (62.50) 7 (70.00) 10 (47.62) 9 (52.94) 7 (48.18) 4 (40.00)

Normal ALT 18 (48.65) 3 (37.50) 3 (30.00) 11 (52.38) 8 (47.06) 10 (58.82) 6 (60.00)

Correlation between Viral load and ALT

0 1-10^3 10^4 10^5 10^6 10^7 10^80.00

10.00

20.00

30.00

40.00

50.00

60.00

70.00

51.35

62.5070.00

47.6252.94

41.18 40.00

48.65

37.5030.00

52.3847.06

58.82 60.00

Raised ALT Normal ALT

p=0.792

Correlation between Viral load and AST

Viral Load 0 1-10^3 10^4 10^5 10^6 10^7 10^8

Raised AST 20 (54.05) 5 (62.50) 8 (80.00) 11 (52.38) 10 (58.82) 10 (58.82) 4 (40.00)

Normal AST 17 (45.95) 3 (37.50) 2 (20.00) 10 (47.62) 7 (41.18) 7 (41.18) 6 (60.00)

0 1-10^3 10^4 10^5 10^6 10^7 10^80.00

10.00

20.00

30.00

40.00

50.00

60.00

70.00

80.00

54.0562.50

80.00

52.3858.82 58.82

40.0045.95

37.50

20.00

47.6241.18 41.18

60.00

Raised AST Normal AST

p=0.706

Prevalence of HCV genotype Genotype No. of subjects (%)

1a 4 (3.33)1b 3 (2.5)2 1 (0.83)3 103 (85.83)4 1 (0.83)

5 or 6 8 (0.67)

1a 1b 2 3 4 5 or 6 0

10

20

30

40

50

60

70

80

90

3.33 2.50.83000000000

0001

85.83

0.830000000000001

6.67

No. of subjects (%)

Summary

1.There is no significant correlation between Anti-HCV to HCV RNA (P=0.692).

As a result since Anti-HCV negativity may be detected in viremic patients molecular methods should be applied specially for suspected cases.

Conclusion