A rapid, low-cost quantitative diagnostic method for hepatitis C virus infection using capillary zone electrophoresis

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 °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.

Microscale electrokinetic‐based analysis of intact cells and viruses

“…These findings allowed to optimize the rapid MECK analysis of these five phages species, in whole blood and plasma samples. A similar study, with clinical relevance, was reported by Attallah et al [60] with the CZE detection and diagnostic of hepatitis C virus in less than 15 min.…”

Capillary electrophoresis based on nucleic acid detection for diagnosing human infectious disease

“…CE detection of microorganisms can be based on the detection of cells [10][11][12][13][14][15][16][17][18], proteins [19][20][21], liposomes [22], or nucleic acids. Among them, nucleic acid detection eliminates many types of false positive and negative results, offering a more precise diagnosis.…”

MS2 and Qβ bacteriophages reveal the contribution of surface hydrophobicity on the mobility of non‐enveloped icosahedral viruses in SDS‐based capillary zone electrophoresis

“…Consequently, non‐enveloped icosahedral viruses possess a global negative surface charge at pH higher than the isoelectric point contributing to proffer them an electrophoretic mobility . Exploiting this characteristic, it has been shown that capillary zone electrophoresis (CZE), a separative analytical technique based on the difference of the electrophoretic mobility of the analytes , can be applied in the separation, detection and quantification of viral, subviral and virus‐like particles in aqueous media . However, in practice, the CZE analysis of viruses is harder than of molecules, where uncontrolled viral particles aggregation and capillary wall adsorption responsible of irreproducible results having been often encountered .…”