Abstract
Background: An estimated 325 million people worldwide live with hepatitis B and/or C and approximately, 5 million people are affected with hepatitis B. in Pakistan.This study aimed at developing PreS protein from Hepatitis B Virus Pakistani isolate (SBS001) with enhanced sensitivity to detect antibodies in serum as a diagnostic method.
 Methods: Gene encoding PreS region from hepatitis B Virus was cloned and expressed in Escherchia coli. The recombinant protein preS-His was purified by Ni-IDA affinity chromatography. Antibodies were raised in rabbit. This protein was screened for detection of antibodies in HBV patients’ sera through ELISA. This ELISA procedure was compared with commercially available Kit used for diagnosis of HBV infection.
 Results: Single band purified recombinant PreS protein was obtained with high titer of antibodies raised in rabbits. This recombinant protein was used in ELISA as antigen coated on the plate. That efficiently detected antibodies present in HBV patients. It was concluded that preS-His antigen/ protein A HRP-conjugate ELISA method was more sensitive than the commercial kit for detecting the antibodies present in HBV patient sera.
 Conclusion: It was concluded that SBS001 PreS recombinant protein can be used in ELISA kits for detection of HBV in Pakistani population.
Highlights
An estimated 325 million people worldwide live with hepatitis B and/or C and approximately, 5 million people are affected with hepatitis B. in Pakistan.This study aimed at developing PreS protein from Hepatitis B Virus Pakistani isolate (SBS001) with enhanced sensitivity to detect antibodies in serum as a diagnostic method
PreS protein with His6 tag attached at Cterminus was successfully cloned in pET 21a (+) vector and expressed in E
The preS domain of hepatitis B virus possesses important biological functions, which are necessary for its infection
Summary
An estimated 325 million people worldwide live with hepatitis B and/or C and approximately, 5 million people are affected with hepatitis B. in Pakistan.This study aimed at developing PreS protein from Hepatitis B Virus Pakistani isolate (SBS001) with enhanced sensitivity to detect antibodies in serum as a diagnostic method. This protein was screened for detection of antibodies in HBV patients’ sera through ELISA. This ELISA procedure was compared with commercially available Kit used for diagnosis of HBV infection. Results: Single band purified recombinant PreS protein was obtained with high titer of antibodies raised in rabbits. This recombinant protein was used in ELISA as antigen coated on the plate. It was concluded that preS-His antigen/ protein A HRP-conjugate ELISA method was more sensitive than the commercial kit for detecting the antibodies present in HBV patient sera. Conclusion: It was concluded that SBS001 PreS recombinant protein can be used in ELISA kits for detection of HBV in Pakistani population
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
