Establishment and performance evaluation of subgenomic RNA detection methods for the 2019 novel coro-navirus
10.3969/j.issn.1006-5725.2024.12.021
- VernacularTitle:新型冠状病毒亚基因组RNA检测方法的建立及性能评估
- Author:
Zhiwei ZHAO
1
;
Cha CHEN
;
Bin HUANG
Author Information
1. 中山大学附属第一医院医学检验科(广州 510080)
- Keywords:
2019 novel coronavirus;
subgenomic RNA;
polymerase chain reaction;
reverse tran-scription
- From:
The Journal of Practical Medicine
2024;40(12):1737-1743
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish a method for detecting the 2019 novel coronavirus subgenomic RNA(sgRNA),and evaluate the performance of the established method.Methods Primers and probes were designed according to the subgenomic sequence of the 2019 novel coronavirus,and a reverse transcription PCR method for sgRNA detection was established.The established method was optimized,including the concentration and propor-tion of primers and probes,extension temperature,reaction volume and template amount.The performance of the method was evaluated,including the limit of detection,sensitivity,specificity and repeatability.sgRNA and genomic RNA(gRNA)were detected in clinical samples,and the results were analyzed.Results In this study,an RT-PCR method for the detection of sgRNA of the 2019 novel coronavirus was established.The limit of detection of this method was 100 copies/mL,and the detection results of common pathogens were negative.The CV of sgRNA in high,medium and low concentration samples were less than 5%.sgRNA was positive in 115 suspected 2019 novel coronavirus infected patients(115/330,34.85%).When the Ct value of gRNA-N was less than 30,the positive rate of sgRNA was 100.00%.When the Ct value of gRNA-N was in the range of 30-32,the positive rate of sgRNA was 68.75%.When the Ct value of gRNA-N was in the range of 32-35,the positive rate of sgRNA was 44.44%.When the Ct value of gRNA-N was greater than 35,the sgRNA was negative.Conclusion The RT-PCR method for the detection of sgRNA of the 2019 novel coronavirus was established,and the detection method was sensitive,specific and reproducible.
