A Rapid On-Site Assay for the Detection of Influenza A by Capillary Convective PCR

Molecular Diagnosis & Therapy
Zhihao ZhuoNingshao Xia

Abstract

Morbidity and mortality from influenza A (Flu A) have increased in recent years. Timely diagnosis and management are critical for disease control. Therefore, the development of a rapid, accurate, and portable analytical method for on-site analysis is imperative. The aim of this work was to develop a rapid, on-site, automated assay for the detection of Flu A and to evaluate the assay. A handheld instrument (TD-01) based on capillary convective polymerase chain reaction (PCR) was developed for rapid on-site detection of Flu A. Since a previous version of the instrument, an automated motion mechanism has been introduced to TD-01 to achieve RNA automated testing. The primers and probe used for Flu A detection were designed according to the Flu A gene sequence of matrix proteins. Finally, we evaluated the detection spectra, sensitivity, specificity, and diagnostic performance of the assay. The TD-01 was able to successfully automatically detect Flu A RNA within 30 min. Results for serially diluted viruses indicated that the lower limit of detection for Flu A was 0.1 TCID50/ml (50% tissue culture infective dose). After evaluating known virus stocks, including 15 strains of Flu A, four strains of Flu B, and two strains of respiratory ...Continue Reading

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Citations

Jun 25, 2019·BioTechniques·Shiyin ZhangNingshao Xia
May 16, 2020·Frontiers in Cellular and Infection Microbiology·Philipp P NelsonChrysanthi Skevaki
Oct 16, 2020·Clinica Chimica Acta; International Journal of Clinical Chemistry·Ya ZhangNingshao Xia

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Datasets Mentioned

BETA
HE818460.1

Methods Mentioned

BETA
chips
reverse transcription PCR
PCR
nucleic acid extraction
electrophoresis

Software Mentioned

DNASTAR
BLAST

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