Development and validation of a simple and rapid method for hepatitis C virus genotyping based on one‑step RT‑qPCR

  • Authors:
    • Xinyun Yang
    • Ting Ding
    • Haifeng Huang
    • Yang Xu
    • Jian Yu
    • Zhanguo Chen
  • View Affiliations

  • Published online on: June 19, 2020     https://doi.org/10.3892/etm.2020.8912
  • Pages: 2284-2290
Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )
Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )


Abstract

Hepatitis C virus (HCV) infections caused by different subtypes require different treatments; therefore, rapid and cost‑effective genotyping methods for the diagnosis of HCV are greatly needed. In the present study, a new method to diagnose HCV subtypes that depends on a one‑step quantitative reverse transcription PCR (RT‑qPCR) and TaqMan fluorescence probe technique is described. Five pairs of primers and five probes were designed, which were able to detect five genotypes in three reaction tubes. One reaction was used to detect the 1b subtype, one was used to detect the 2a and 6a subtypes, and the other was used to detect the 3a and 3b subtypes. Rigorous performance validation was implemented for five aspects: Precision, sensitivity, accuracy, specificity and anti‑interference. The HCV subtype that infected 289 patients was evaluated in the present study via RT‑qPCR and verified by sequencing. The results revealed that the 1b subtype accounted for 45% of infections, the 2a subtype accounted for 9% of infections, the 3a subtype accounted for 13% of infections, the 3b subtype accounted for 18% of infections, and the 6a subtype accounted for 15% of infections. The analytical sensitivity for the detection of each of the five HCV subtypes was 1,000 IU/ml. The new method performed well in the performance validation mentioned above, indicating its effectiveness as a HCV genotyping method. RT‑qPCR has mitigated some of the former challenges of existing HCV genotyping methods, including the time commitment, expense, and inaccuracy of such methods. The performance validation of this new method showed that RT‑qPCR is reliable enough to be widely applied in China for HCV genotyping.

Related Articles

Journal Cover

September-2020
Volume 20 Issue 3

Print ISSN: 1792-0981
Online ISSN:1792-1015

Sign up for eToc alerts

Recommend to Library

Copy and paste a formatted citation
x
Spandidos Publications style
Yang X, Ding T, Huang H, Xu Y, Yu J and Chen Z: Development and validation of a simple and rapid method for hepatitis C virus genotyping based on one‑step RT‑qPCR. Exp Ther Med 20: 2284-2290, 2020
APA
Yang, X., Ding, T., Huang, H., Xu, Y., Yu, J., & Chen, Z. (2020). Development and validation of a simple and rapid method for hepatitis C virus genotyping based on one‑step RT‑qPCR. Experimental and Therapeutic Medicine, 20, 2284-2290. https://doi.org/10.3892/etm.2020.8912
MLA
Yang, X., Ding, T., Huang, H., Xu, Y., Yu, J., Chen, Z."Development and validation of a simple and rapid method for hepatitis C virus genotyping based on one‑step RT‑qPCR". Experimental and Therapeutic Medicine 20.3 (2020): 2284-2290.
Chicago
Yang, X., Ding, T., Huang, H., Xu, Y., Yu, J., Chen, Z."Development and validation of a simple and rapid method for hepatitis C virus genotyping based on one‑step RT‑qPCR". Experimental and Therapeutic Medicine 20, no. 3 (2020): 2284-2290. https://doi.org/10.3892/etm.2020.8912