Comparison of the diagnostic value of one-step and two-step real-time Reverse transcription polymerase chain reaction (Real time RT-PCR) to detection of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2)
کد: G-31064
نویسندگان: Asra Malekshahi, Sayyad Khanizadeh ©, Gholamreza Goudarzi, Gholamreza Talei, Sara Gravand, Mohsen Hemati Dinarvand
زمان بندی: زمان بندی نشده!
برچسب: ویروس شناسی پزشکی
دانلود: دانلود پوستر
خلاصه مقاله:
Background and Aim
The gold standard test for the diagnosis of SARS-CoV-2 is the one-step RT-qPCR method, which requires expensive probes and one-step reagents and not always available.
Methods
In the current study, a cost-effective technique based on SYBR green to detection of SARS‑CoV‑2 was evaluated. So that the specific primers for S and N genes were designed for use in the SYBR green-based two-step RT-qPCR method and then, the cross-reactivity test was performed with positive samples for other coronavirus and respiratory viruses. Moreover, the analytical sensitivity test was carried out with 8 dilutions (10-fold) of SARS-CoV-2 for one-step and two-step RT-qPCR. Lastly, the SARS-CoV-2 clinical samples (n= 210) were tested by these two methods and Receiver operating characteristic (ROC) analysis was performed to investigate incremental diagnostic value of each genes in the study methods
Results
The two-step RT-qPCR did not show any amplification of the positive samples of other respiratory viruses; it was able to detect until 7th dilutions of the SARS-CoV-2 sample. Furthermore, two-step qPCR detected 90% of the positive samples correctly. ROC analysis revealed a diagnostic ability of two-step RT-qPCR method for SARS-CoV-2 with an area under ROC curve of ≥0.7 (p ˂ 0.05) and relatively high sensitivity and specificity. Combination of N and S genes increased the sensitivity up to 88%, specificity up to 86% and area under the ROC curve up to 0.85 (95% confidence interval (95% CI): 0.72 to 0.93, p=0.0461)
Conclusion
Our findings indicated that the two-step method has comparable sensitivity and specificity to the one-step method. Therefore, this method can be considered as alternative diagnostic method for diagnosis and monitoring of COVID-19 patients.
Keywords
SARS-CoV-2; RT-qPCR; SYBR Green; TaqMan
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