Evaluation of Seven Rapid Diagnostic Tests for Detection of Hepatitis C Virus Antibodies in China

Bing Chen; Zhong-Hui Ma; Bing Xu; Hao Chang; Xiao-Xia He; Li-Jian Pei; Ya-Nan Ren; Wen-Ge Xing


J Viral Hepat. 2021;28(4):657-663. 

In This Article

Materials and Methods

Samples and Kits

Serum samples were collected from intravenous drug users and blood donors in Yunnan Province and Gansu Province in China. The selected serum samples were diagnosed as HCV negative or positive by ELISA reagents (Murex, DiaSorin; Wantai BioPharm), RIBA reagents (Wantai BioPharm), and real-time PCR reagents (Cobas, Roche). The result judgement referred to per manufacturer's instructions of the testing reagents.

In this study, Seven HCV antibody RDT kits approved by CFDA were selected for performance evaluation, and the manufactures of HCV RDT kits include Kehua Bio-Engineering, Wantai BioPharm, Coretests, InTec Products, Newscan Coast, Wondfo, Biotest, which were represented by A, B, C, D, E, F, G, respectively. The detailed comparisons of selected HCV RDT kits were shown in supplementary material Table S1.

Evaluation Procedures

Before the evaluation of the experiment, all the operators were trained in performing and/or interpreting the HCV RDT assays. All specimens in all HCV serum panels were tested by the selected seven HCV RDT kits, and all assays were performed by one operator who was blinded to the reference results according to per manufacturer's instructions manuals. In addition, the results were visually interpreted by two others independently readers, and the readers were blinded to the reference results as well as the other's reading. Then, the results of different HCV RDT kits were recorded, compared and analysed with reference results in HCV serum panels.

Construction of HCV Serum Panel

HCV Basic Panel. HCV basic panel was mainly used to evaluate the clinical sensitivity, clinical specificity, positive predictive value (PPV) and negative predictive value (NPV) of HCV RDT kits. And the basic panel consisted of 100 serum samples, including 50 HCV antibody-positive samples and 50 HCV-negative samples.

HCV Seroconversion Panel. HCV seroconversion panel was a series of samples, continuously collected over a period of time, from a HCV infected individual during the period between primary infection and HCV antibody production. And it was used to evaluate the sensitivity of HCV RDT kits in early infection of HCV. A total of 40 samples from four seroconversion panels (PHV906, PHV908, PHV914, PHV921; Seracare Life Sciences) were tested on each of the seven HCV RDT kits evaluated.

HCV Analytical Specificity Panel. HCV analytical specificity panel was used to evaluate the analytical specificity of HCV RDT kits. This set of serum panel consisted of 45 HCV antibody-negative samples, but these samples have multiple potentially interference factors in immunoassays. For example, these samples had haemoglobin, triglycerides, and other viral infections, such as HIV, HBV and syphilis antibody positive.

HCV Mixed Titre Performance Panel. HCV mixed titre performance panel was used to evaluate the different antibodies titres of HCV RDT kits. This serum panel was composed of 16 undiluted and naturally occurring serum samples, which had antibodies reactivity ranging from negative to strongly positive for anti-HCV. One negative serum has been included as nonreactive controls.

HCV Genotype Qualification Panel. HCV genotype qualification panel was used to evaluate the ability of different genotypes detection of HCV RDT kits. Commercially HCV genotype qualification panel (Panel: 2400–0182) was purchased from Seracare Company, which consists of eight HCV RNA positive and one HCV RNA negative. Single-positive samples from different infected patients. HCV genotypes (subtypes) of the eight HCV RNA-positive samples were HCV 1a, 1b, 2a, 2b, 3a, 4a, 5a, 6, respectively.

HCV Characteristic Panel. HCV characteristic panel was used to evaluate the ability of different bands detection of HCV RDT kits. And this serum panel was composed of 20 serum samples, which had a single band, two bands, three bands, four bands, and whole bands, respectively. The background information of HCV characteristic panel shows in Table S2.

Statistical Analysis

Data analyses were performed using GraphPad PRISM 8.0. Clinical sensitivity and clinical specificity with confidence intervals (CIs) for each HCV RDT kit were calculated by comparing results obtained using the RDT with the reference result.