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• W2904853651 abstract "Hepatitis C (HCV) and Hepatitis B virus (HBV) infections are major public healthissue worldwide. These two hepatotropic viruses share same ways of transmissionand also coinfection with these two viruses is not unusual, especially in areas with ahigh prevalence of HBV infection and among people at high risk for parenteralinfection (Liu and Hou 2006). It is imperative that effective and reliable techniquesfor hepatitis C virus (HCV) and hepatitis B (HBV) infection diagnosis be devised.Additionally, non-reactive and misdiagnosis, often demanding iteration of tests forconfirmation. Biosensors based on surface plasmon resonance (SPR) detection assayprovides a solution for these limitations. Additionally, the SPR can overcome thecontamination susceptibility of molecular methods that rely heavily on the purity ofthe template nucleic acid and that would result in false positives.This study proposes an optimized SPR protocol for large-scale Hepatitis C and Bsamples screening. HCV and HBV detection chips were set up separately. The HCVdetection chip was established by immobilization of HCV Core genotype 1, HCVCore genotype 3a and HCV NS5 genotype 3a antigens for hepatitis C genotypescreening. On the other hand, HBs Ag and HBsAg antibody were used to establishthe HBV screening chip. The limit of detection (LOD) was calculated for eachimmobilized flow cell of each established chip used 20 human donor serums wereeach spiked with HCV antibody for HCV established chip and 20 human donorserums were each spiked with HBsAg antibody for HBV established chip. LOD ofHCV detection chip was in the range of 14.4 – 33.48 pg/ ml followed by a range of18- 25.2 pg/ml for HBV detection chip used .A total 137 HCV positive and negative samples were collected. All of the 137 testedas positive or negative HCV samples, were analyzed for genotype 1 and genotype 3awith established HCV screening chip. 37 samples were tested positive for HCVantibody, showed 100% positivity for genotype 1. Consequently, 6.6 % testedpositive for core genotype 3a, while 16% tested positive for NS5 genotype 3a.Among the genotype 3a samples tested positive, only one sample showed positiveresults for both core 3a and NS5 3a.A total 400 HBV positive and negative samples were collected. All 100 positiveHBsAg, 100 negative HBsAg and 200 negative HBsAg antibody samples wereconfirmed serologically by the HBV established chips.Finally, the HBsAg, HCV core genotype 1 and HBsAg antibody were immobilizedto establish the dual detection chip. Based on the international distribution of HCVgenotype 1, it was chosen to establish the dual detection chip. The LOD for this dualdetection chip was in the range of 12.6–26.4 pg/ml. To determine the LOD for thisestablished chip, 20 human donor serums were used and each sample spiked withHCV antibody and HBsAg antibody individualy and analysed with established chip.Out of total 137 samples, 37 samples tested positive for HCV antibody and 100tested negative for HCV antibody. Among oof 400 samples, 100 samples testedpositive for HBsAg, 100 samples tested negative for HBsAg and 200 samples testedHBsAg antibody negative.A comparison was made between the SPR and ELISA techniques in the diagnosis ofHCV and HBV infection. It was found that there was a strong correlation betweenSPR and ELISA test when used to detect HCV and HBV in the samples. Thecorrelation coefficient obtained with the two techniques approached 1 (P< 0.01).The range of linear dose was observed for each immobilized flow cell with acoefficient of determination between 98 to 99%.The novelty of this study is the ability to serologically detect HCV and HBVantigens and antibody, as well as HCV genotypes mixture simultaneously. LOD fordeveloped SPR chips (observed between 14.4 – 33.48 pg/ml) showed higherefficiency when compared to HCV and HBV commercially used enzyme-linkedimmunosorbent assay (ELISA) and chemiluminescent immunoassay (ChLIA)(ng/ml). Due to the low detection limit of SPR compared to ELISA and ChLIA, itcan detect false negative samples caused by lower level of antibody and antigen thanELISA and ChLIA detection limit, a greater efficiency lacking in the later mentionedapplication.In conclusion, the optimized SPR approach can serve as a standard operatingprocedure (SOP) for national blood screening centers. Taken together, data indicatethat the assays developed are highly reproducible, specific and sensitive , accuracy,precision, repeatability, linearity, range and robustness.. This biosensor-based assayis a more efficient tool for accurate screening of antibody and antigen in HCV andHBV infected patient serum, while retaining the advantages of ELISA. Moreover,the inbuilt robotic automated system with reusable chip is on the top of novelty forthis dual antigen and antibody detection assay for suspected co-infected samples." @default.
• W2904853651 created "2018-12-22" @default.
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• W2904853651 date "2015-05-01" @default.
• W2904853651 modified "2023-09-27" @default.
• W2904853651 title "Development of surface plasmon resonance based assay for simultaneous detection of Hepatitis C and B viral infections" @default.
• W2904853651 hasPublicationYear "2015" @default.
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