A rapid, low-cost quantitative diagnostic method for hepatitis C virus infection using capillary zone electrophoresis
EUROPEAN JOURNAL OF CLINICAL MICROBIOLOGY & INFECTIOUS DISEASES
Authors: Attallah, A. M.; Abdallah, S. O.; El-desouky, M. A.; El-Far, M.; Omran, M. M.; Farid, K.; Abdelrazek, M. A.; Shabaka, M. N.; Zaghloul, H.; Fawzy, A. M.; Bazeed, F. B.
Abstract
Hepatitis C virus (HCV)-RNA amplification is a costly procedure in terms of time and reagents. Consequently, the search for more a cost-effective specific HCV diagnostic method is of great interest. Capillary zone electrophoresis (CZE) methods that detect HCV in serum, plasma, whole blood, and ascites without the need for sample pretreatment are not currently available. Here, a CZE method was developed that detects a larger specific peak in serum and other body fluids of HCV-infected patients than that found in healthy or hepatitis B virus (HBV)-infected individuals. The nature of the HCV peak was investigated using biochemical treatments, including RNase, DNase, and chymotrypsin enzymes. Electroeluted HCV peak was applied to transmission electron microscopy; electron micrographs showed that the HCV peak was attributed to virus-like particles with diameter and morphological properties similar to non-enveloped HCV nucleocapsids. The determination of CZE-HCV and HCV-RNA levels using quantitative real-time reverse transcriptase-polymerase chain reaction (qRT-PCR) in 258 subjects revealed that these two tests were highly correlated (r = 0.92, p < 0.0001). One important issue of HCV testing is the storage conditions of serum to obtain reliable results. Serum samples at -20 A degrees C showed the best preservation of the HCV peak up to one year. In conclusion, we detected HCV using CZE in a microliters volume from different body fluids. Besides the stability of samples in maintaining their peak height, the HCV-CZE test is rapid (< 15 min) and a well-suited and low-cost technique. Thus, a major improvement in the quantitative diagnosis of HCV infection was established.
Hepatitis C point-of-care diagnostics: in search of a single visit diagnosis
EXPERT REVIEW OF MOLECULAR DIAGNOSTICS
Authors: Grebely, Jason; Applegate, Tanya L.; Cunningham, Philip; Feld, Jordan J.
Abstract
Introduction: The availability of simple, tolerable, therapies for hepatitis C virus (HCV) infection with responses > 95% is one of the greatest medical advances in decades, offering an opportunity to reverse the rising burden due to HCV and strive towards HCV elimination. A key challenge moving forward will be to ensure that those who are undiagnosed are made aware of their infection, receive HCV therapy and achieve viral cure. The availability of point-of-care tests for HCV infection has the potential to simplify testing algorithms, increase diagnoses, and facilitate linkage to treatment. Areas covered: This commentary explores why point-of-care tests for HCV are needed, what markers of HCV can be measured, methods for sample collection, where HCV testing can occur, and the remaining challenges for HCV point-of-care testing. Expert commentary: Currently, we have reached an era where there are now several commercial assays to detect HCV RNA (active HCV infection) in 60-90 min, and have reached a single visit HCV diagnosis. In the future, it is hoped that further technological advances will enable access to low-cost, rapid, and accurate assays for HCV RNA detection, improving the number of people diagnosed with HCV infection and contributing to global elimination efforts.