HCV Sample Dilution And Inactivation Lab Report
II. HCV Sample Dilution and Inactivation
From each sample 225 µL was transferred to a microcentrifuge tube, which containing 25 µL of Triton X-100 Solution; the tubes were Incubated 30 minutes at 37ºC, and then the samples were incubated at 56ºC for 30 min in water path (for dissotiation of HCV core antigen-antibody complex).
Assay procedure
1. All reagents were leaved to reach room temperature before use, and then shacked thoroughly with no foam.
2. The number of strips required for the test was fixed on a plate holder.
3. Sample inactivation (dissotiation of HCV core antigen-antibody complex step).
4. 100 µL of inactivated sample and HCVcAg standard were added to Anti-HCVcAg Antibody Coated Plate.
5. The Plate was Covered and incubated …show more content…
The plate was washing by aspiration of liquid from each well then 250 µL of washing solution was added to each well. The wells were allowed to soak for 20-30 seconds then the content of each well was aspirated again.
7. The repeated washing step was repeated for 5 times. After the final wash cycle, the plate was turned into blotting paper or clean towel and tapped to remove any remainders. The correct washing procedure is critical for successful performance of the test. Also, the well should not be allowed to dry during the assay procedure.
8. Immediately after wash, 100 µL was taken from the diluted FITC-conjugated Anti-HCVcore antigen monoclonal antibody was added to every well.
9. Then plate was covered and incubated at room temperature for 1 hour on an orbital shaker. After the end of the incubation, the plate cover was removed and discarded and washing was done as mentioned …show more content…
Means a strong negative linear relationship.
–0.50. Means a moderate negative relationship.
–0.30. Means a weak negative linear relationship.
No linear relationship
0. No linear relationship.
Positive linear relationship
+0.30. Means a weak positive linear relationship.
+0.50. Means a moderate positive relationship.
+0.70. Means a strong positive linear relationship.
Exactly +1. Means a perfect positive linear relationship.
Discussion
Hepatitis C virus infection is a global health threat worldwide approximately 3% of worldwide population is infected with HCV, and more than 350.000 die every year from hepatitis C related liver damage. The prevalence of HCV infection varies throughout the world. Egypt has the highest prevelance of adult HCV infection in the world. HCV transmission is ongoing in Egypt (165.000 new infections annually). There are at least 5,459,000 infected with HCV who are infectious to others, which is the major reservoir of infection with HCV in the world.
Anti-HCV assays are the most common serological markers for the diagnosis of HCV infection and are used as the first-line screening test in the community. However, most anti-HCV assays do not correlate well with HCV
From each sample 225 µL was transferred to a microcentrifuge tube, which containing 25 µL of Triton X-100 Solution; the tubes were Incubated 30 minutes at 37ºC, and then the samples were incubated at 56ºC for 30 min in water path (for dissotiation of HCV core antigen-antibody complex).
Assay procedure
1. All reagents were leaved to reach room temperature before use, and then shacked thoroughly with no foam.
2. The number of strips required for the test was fixed on a plate holder.
3. Sample inactivation (dissotiation of HCV core antigen-antibody complex step).
4. 100 µL of inactivated sample and HCVcAg standard were added to Anti-HCVcAg Antibody Coated Plate.
5. The Plate was Covered and incubated …show more content…
The plate was washing by aspiration of liquid from each well then 250 µL of washing solution was added to each well. The wells were allowed to soak for 20-30 seconds then the content of each well was aspirated again.
7. The repeated washing step was repeated for 5 times. After the final wash cycle, the plate was turned into blotting paper or clean towel and tapped to remove any remainders. The correct washing procedure is critical for successful performance of the test. Also, the well should not be allowed to dry during the assay procedure.
8. Immediately after wash, 100 µL was taken from the diluted FITC-conjugated Anti-HCVcore antigen monoclonal antibody was added to every well.
9. Then plate was covered and incubated at room temperature for 1 hour on an orbital shaker. After the end of the incubation, the plate cover was removed and discarded and washing was done as mentioned …show more content…
Means a strong negative linear relationship.
–0.50. Means a moderate negative relationship.
–0.30. Means a weak negative linear relationship.
No linear relationship
0. No linear relationship.
Positive linear relationship
+0.30. Means a weak positive linear relationship.
+0.50. Means a moderate positive relationship.
+0.70. Means a strong positive linear relationship.
Exactly +1. Means a perfect positive linear relationship.
Discussion
Hepatitis C virus infection is a global health threat worldwide approximately 3% of worldwide population is infected with HCV, and more than 350.000 die every year from hepatitis C related liver damage. The prevalence of HCV infection varies throughout the world. Egypt has the highest prevelance of adult HCV infection in the world. HCV transmission is ongoing in Egypt (165.000 new infections annually). There are at least 5,459,000 infected with HCV who are infectious to others, which is the major reservoir of infection with HCV in the world.
Anti-HCV assays are the most common serological markers for the diagnosis of HCV infection and are used as the first-line screening test in the community. However, most anti-HCV assays do not correlate well with HCV