Development and application of a rapid scheme for detection of respiratory virus nucleic acid.

Yuanyuan Huang; Yu Wang; Chengxing Zhou; Zhichao Zhou; Bingliang Zhou; Wenkuan Liu; Rong Zhou; Hong Cao

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Development and application of a rapid scheme for detection of respiratory virus nucleic acid.

Author: Yuanyuan HUANG ¹ ; Yu WANG ² ; Chengxing ZHOU ¹ ; Zhichao ZHOU ³ ; Bingliang ZHOU ¹ ; Wenkuan LIU ⁴ ; Rong ZHOU ² ; Hong CAO ¹
Author Information

1. Guangdong Provincial Key Laboratory of Tropical Disease Research, Department of Microbiology, School of Public Health, Southern Medical University, Guangzhou 510515, Guangdong, China.
2. Guangzhou Laboratory, Guangzhou 510005, Guangdong, China.
3. Department of Radiation Medicine, School of Public Health, Southern Medical University, Guangzhou 510515, Guangdong, China.
4. State Key Laboratory of Respiratory Disease, The First Affiliated Hospital of Guangzhou Medical University, Guangzhou Medical University, Guangzhou 510120, Guangdong, China.
Publication Type:Journal Article
Keywords: extraction-free respiratory virus treatment reagent (RTU); quantitative real-time polymerase chain reaction; rapid detection; respiratory virus
MeSH: Humans; Respiratory Syncytial Virus Infections/diagnosis*; Respiratory Syncytial Virus, Human/genetics*; Nucleic Acid Amplification Techniques; Real-Time Polymerase Chain Reaction; Adenoviridae; Sensitivity and Specificity
From: Chinese Journal of Biotechnology 2023;39(9):3838-3848
CountryChina
Language:Chinese
Abstract: This study aimed to develop a portable, accurate and easy-to-operate scheme for rapid detection of respiratory virus nucleic acid. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to verify the effect of extraction-free respiratory virus treatment reagent (RTU) on viral nucleic acid treatment and the effect of ultra-fast fluorescence quantitative PCR instrument (FQ-8A) on nucleic acid amplification, respectively. RTU and FQ-8A were combined to develop a rapid detection scheme for respiratory virus nucleic acid, and the positive detection rate was judged by C_t value using a fluorescence quantitative PCR instrument, and the accuracy of the scheme in clinical samples detection was investigated. The results showed that RTU had comparable sensitivity to the automatic nucleic acid extraction instrument, its extraction efficiency was comparable to the other 3 extraction methods when extracting samples of different virus types, but the extraction time of RTU was less than 5 min. FQ-8A had good consistency in detection respiratory syncytial virus (RSV) and adenovirus (ADV) compared with the control instrument ABI-7500, with kappa coefficients of 0.938 (P < 0.001) and 0.887 (P < 0.001), respectively, but the amplification time was only about 0.5 h. The RTU and FQ-8A combined rapid detection scheme had a highly consistent detection rate with the conventional detection scheme, with a sensitivity of 91.70% and specificity of 100%, and a kappa coefficient was 0.944 (P < 0.001). In conclusion, by combining RTU with FQ-8A, a rapid respiratory virus nucleic acid detection scheme was developed, the whole process could be completed in 35 min. The scheme is accurate and easy-to-operate, and can provide important support for the rapid diagnosis and treatment of respiratory virus.