Hepatitis C virus (HCV) do harm to people's health. The present study aims to establish a simple HCV detection method by reverse transcription-loop mediated isothermal amplification technique (RT-LAMP). A total of 75 clinical samples were collected and pre-detected by fluorescence quantitative-polymerase chain reaction (FQ-PCR), which was considered as the gold standard. Firstly, four common primers were designed according to the conservative 5'UTR region of HCV on the NCBI website to establish an integrated RT-LAMP reaction system. Then, the reaction efficiency was evaluated by adding Taq DNA polymerase to the conventional system. The specificity of RT-LAMP was evaluated by observing the length of fragment after endonuclease digestion and by a templates exchange assay, the sensitivity of RT-LAMP was evaluated by detection of diluted clinical templates. The results were compared with that of reverse transcription polymerase chain reaction (RT-PCR). At the same time, the performance was judged using calcein and hydroxynaphthol blue (HNB) stain methods, The two results were compared with that of electrophoresis method. At last, 75 clinical samples were detected by both RT-LAMP and RT-PCR methods. Results showed that the reaction efficiency was increased 20 minutes after adding Taq DNA polymerase to the normal RT-LAMP system. RT-LAMP showed good specificity, the digestion length was consistent with our expectation (216 bp) after restriction endonuclease cleavage assay, and only the templates of HCV were amplified using the common RT-LAMP primers. After detection of diluted temples, the sensitivity of RT-LAMP was 10 IU/tube, which was 10 fold higher than that of PCR. In addition, the results using calcein and HNB stain methods were the same with that of electrophoresis method. After detection of all 75 clinical samples, the results indicated that RT-LAMP had worse consistency with RT-PCR (P < 0.05, Kappa=0.375). However, RT-LAMP, on the contrary, showed good consistency with FQ-PCR (P > 0.05, Kappa=0.762). In conclusion, RT-LAMP has characteristics of simplicity, specificity and sensitivity, and this technique is suitable for the primary care hospitals.
ObjectiveTo systematically review the diagnostic value of Antibodies to hepatitis C virus (anti-HCV) in serum tested by Chemiluminescence Assay for patients with hepatitis C. MethodsSuch databases as PubMed, EMbase, The Cochrane Library (Issue 1, 2014), CBM, CNKI, VIP, and WanFang Data were searched from the date of their establishment to March 1st, 2014, and the relevant journals were also manually searched to collect the studies which evaluated the diagnostic value of anti-HCV in serum tested by chemiluminescence assay for patients with hepatitis C. Literature screening according to the inclusion and exclusion criteria, data extraction and methodological quality assessment were completed by two reviewers independently. Meta-analysis was then conducted using MetaDisc 1.4 software. ResultsA total of 12 studies involving 1 252 patients were enrolled. The results of meta-analysis indicated that, the pooled sensitivity was 0.99 (95%CI 0.98 to 0.99, P=0.000 1), and the pooled specificity was 0.98 (95%CI 0.98 to 0.99, P=0.000 0). The positive likelihood ratio was 42.53 (95%CI 18.05 to 100.23, P=0.000 0) and the negative likelihood ratio was 0.02 (95%CI 0.01 to 0.03, P=0.026 2). The diagnostic odds ratio was 3 005.82 (95%CI 1 257.08 to 7 187.29, P=0.006), overall area under the curve (AUC) was 0.9 977 (SE=0.000 8), and Q* was 0.983 3 (SE=0.003 6). ConclusionAnti-HCV in serum tested by chemiluminescence assay is sensitive, specific and stable, which is suitable for extensive application in clinics.