A Convenient Nucleic Acid Test on the Basis of the Capillary Convective PCR for the On-Site Detection of Enterovirus 71
| Autor: | Ping-Hei Chen, Pingguo Liu, Ningshao Xia, Shengxiang Ge, Jieyu Chen, Penglin Wang, Jin Wang, Shuizhen He, Yanyan Lin, Xu Feihai, Zhou Wenbin, Miaoge Xue, Shiyin Zhang |
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| Rok vydání: | 2014 |
| Předmět: |
China
Pcr cloning Diagnosis tool Biology Polymerase Chain Reaction Sensitivity and Specificity Pathology and Forensic Medicine law.invention law medicine Enterovirus 71 Humans Child Polymerase chain reaction medicine.diagnostic_test Nucleic acid test Equipment Design Dipstick biology.organism_classification Virology Enterovirus A Human Visual detection Nucleic acid RNA Viral Molecular Medicine Hand Foot and Mouth Disease |
| Zdroj: | The Journal of Molecular Diagnostics. 16:452-458 |
| ISSN: | 1525-1578 |
| Popis: | The recent and continuing epidemic of enterovirus 71 in China has affected millions of children and resulted in thousands of deaths. Timely diagnosis and management is essential for disease control. Current enterovirus 71 molecular tests require resources that are unavailable for on-site testing. We have developed a simple-to-operate nucleic acid test, the convenient and integrated nucleic acid test, for local medical institutions. It uses a convective PCR for rapid amplification, a dipstick for visual detection of PCR products, and a simple commercial kit for nucleic acid extraction. By using a specially designed reagent and reaction tube containing a dipstick, the amplification and detection processes are well integrated and simplified. Moreover, cross contamination that may be caused by an open-tube detection system can be avoided. On the basis of the convenient and integrated nucleic acid test, an enterovirus 71 assay for on-site testing was developed. After evaluating known hand, foot, and mouth disease virus stocks of 17 strains of 11 different serotypes, this assay showed a favorable detection spectrum and no cross-reactivity. Its clinical performance was established by testing 141 clinical samples and comparing the results with a nested RT-PCR method. The assay showed a clinical sensitivity and specificity of 98.5% and 100%, respectively. Our results suggest that this convenient and integrated nucleic acid test enterovirus 71 assay may serve as an on-site diagnosis tool. |
| Databáze: | OpenAIRE |
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