University of Khartoum

Assessment of Three Real-time Polymerase Chain Reaction Assays for Diagnosis of Hepatitis B, C and Human Immunodeficiency Virus Type-1 among Blood Donors in Khartoum State-Sudan

Assessment of Three Real-time Polymerase Chain Reaction Assays for Diagnosis of Hepatitis B, C and Human Immunodeficiency Virus Type-1 among Blood Donors in Khartoum State-Sudan

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Title: Assessment of Three Real-time Polymerase Chain Reaction Assays for Diagnosis of Hepatitis B, C and Human Immunodeficiency Virus Type-1 among Blood Donors in Khartoum State-Sudan
Author: MOHAMMED, ELMOEIZ ALI ELNAGI
Abstract: Background: Careful screening of donated blood is mandatory and crucial. The tests that currently used for the detection of HBV, HCV and HIV-1 are primarily serological that use an enzyme-linked immuno-sorbent assay (ELISA). The ELISA test is cost-effective compared to molecular testing as it’s cheaper by 40%, however, it has some disadvantages such as missing of infected individuals during the serological window period and antibody cross-reactivity that may lead to false positive tests. Therefore, molecular techniques are more reliable and sensitive. In addition, molecular techniques have the capability to detect viral nucleic acid which minimizes the danger of transmitting serious diseases through blood transfusion. Objectives: The aim of this study was to assess the efficacy and the analytical performance of three commercial real-time PCR assays for diagnosis and quantification of HBV (DNA), HCV (RNA), and HIV-1 (RNA) in plasma samples from the blood donors of the central blood bank and national health laboratory in Khartoum state. Material and methods: This cross-sectional analytical study was conducted in Elzahrawi medical laboratory and Aliaa specialist hospital laboratory at Khartoum, during the period between 2014 and 2018. A total of 45896 plasma samples from apparently healthy donors were tested serologically by ELISA for the detection of HBV, HCV, and HIV-1. Out of these, 990 blood donors were randomly selected and included in this study. For each of HBV, HCV, and HIV-1, a total of 330 blood donors (including 200 serologically positive blood donors and 130 serologically negative blood donors as control) were tested by three different commercial real-time PCR assays that included genesig® real-time PCR Diagnostic assay, Sansure Quantitative Diagnostic assay, and RealMOD™ Green Real-time PCR Diagnostic assay. The statistical analysis of the data was performed and analyzed by SPSS version 23 using descriptive statistics, linear regression analysis & Pearson correlation coefficient, ANOVA, and post-hoc test. Results: The results of the three real-time PCR assays yielded a broad range of detection for all the three viruses and had specificities of 100%. The analytical sensitivity (the lower limit of detection) of the three real-time PCR assays for HBV DNA used in this study (genesig® real-time PCR advanced detection Kit, Sansure quantitative fluorescence diagnostic Kit, and RealMOD™ Green Real-time PCR diagnostic kit) were 250 copies/ml (56 IU/ml), 500 copies/ml (111 IU/ml), 500 copies/ml (111 IU/ml) respectively. For HCV RNA it was estimated to be <100 copies/ml (<40 IU/ml) for all three assays; and for HIV-1 RNA it was estimated to be 100 copies/ml for all three assays also. These assays showed similar specificity with excellent reproducibility and their results were also in agreement; however, non-significant underestimation of DNA levels could be present in the third assay (RealMOD™ Green Real-time PCR). Conclusion: In conclusion, the three real-time PCR assays gave an excellent agreement for 99.9% of the samples. In addition to that, they demonstrate a very good analytical sensitivity, great reproducibility, 100% specificity, broad linear quantitative range, and proved to be satisfactory for routine laboratory diagnosis of the three viruses (HBV, HCV, and HIV-1).
URI: http://khartoumspace.uofk.edu/123456789/27444


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