A rapid and sensitive recombinase aided amplification assay to detect hepatitis B virus without DNA extraction
Abstract Background Hepatitis B virus (HBV) infection is the major public health problem worldwide. In clinical practice, serological and molecular assays are the most commonly used diagnostic methods to detect HBV infection in clinical practices. Methods Here we present a rapid and sensitive recomb...
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| Format: | Article |
| Language: | English |
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BMC
2019-03-01
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| Series: | BMC Infectious Diseases |
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| Online Access: | http://link.springer.com/article/10.1186/s12879-019-3814-9 |
| _version_ | 1818313523796639744 |
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| author | Xin-xin Shen Fang-zhou Qiu Li-Ping Shen Ten-fei Yan Meng-chuan Zhao Ju-Ju Qi Chen Chen Li Zhao Le Wang Zhi-shan Feng Xue-jun Ma |
| author_facet | Xin-xin Shen Fang-zhou Qiu Li-Ping Shen Ten-fei Yan Meng-chuan Zhao Ju-Ju Qi Chen Chen Li Zhao Le Wang Zhi-shan Feng Xue-jun Ma |
| author_sort | Xin-xin Shen |
| collection | DOAJ |
| description | Abstract Background Hepatitis B virus (HBV) infection is the major public health problem worldwide. In clinical practice, serological and molecular assays are the most commonly used diagnostic methods to detect HBV infection in clinical practices. Methods Here we present a rapid and sensitive recombinase aided amplification assay (RAA) to detect HBV at 39.0 °C for 30 min without DNA extraction from serum samples. The analytical sensitivity of RAA assay was 100 copies per reaction and showed no cross reaction with human immunodeficiency virus (HIV) and hepatitis C virus (HCV). The universality of RAA assay was validated by testing of 41 archived serum samples with predefined HBV genotypes (B, C and D). Results A total of 130 archived suspected HBV infected serum samples were detected by commercial qPCR with DNA extraction and RAA assay without DNA extraction (heat-treatment). Compared with qPCR assay as a reference, the RAA assay obtained 95.7% sensitivity and 100% specificity and a kappa value of 0.818. Conclusions We developed a rapid, convenient, highly sensitive and specific method to detect HBV without DNA extraction in clinical samples. This RAA method of HBV detection is very suitable for clinical testing. |
| first_indexed | 2024-12-13T08:35:06Z |
| format | Article |
| id | doaj.art-a0b1dd95f68a4d38b78a027a4cafd9d1 |
| institution | Directory Open Access Journal |
| issn | 1471-2334 |
| language | English |
| last_indexed | 2024-12-13T08:35:06Z |
| publishDate | 2019-03-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Infectious Diseases |
| spelling | doaj.art-a0b1dd95f68a4d38b78a027a4cafd9d12022-12-21T23:53:39ZengBMCBMC Infectious Diseases1471-23342019-03-011911510.1186/s12879-019-3814-9A rapid and sensitive recombinase aided amplification assay to detect hepatitis B virus without DNA extractionXin-xin Shen0Fang-zhou Qiu1Li-Ping Shen2Ten-fei Yan3Meng-chuan Zhao4Ju-Ju Qi5Chen Chen6Li Zhao7Le Wang8Zhi-shan Feng9Xue-jun Ma10NHC Key Laboratory of Medical Virology and Viral Diseases, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and PreventionNHC Key Laboratory of Medical Virology and Viral Diseases, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and PreventionNHC Key Laboratory of Medical Virology and Viral Diseases, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and PreventionMyasthenia Gravis Research Institute, The First Hospital of ShijiazhuangChildren’s Hospital of Hebei ProvinceNHC Key Laboratory of Medical Virology and Viral Diseases, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and PreventionNHC Key Laboratory of Medical Virology and Viral Diseases, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and PreventionNHC Key Laboratory of Medical Virology and Viral Diseases, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and PreventionChildren’s Hospital of Hebei ProvinceChildren’s Hospital of Hebei ProvinceNHC Key Laboratory of Medical Virology and Viral Diseases, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and PreventionAbstract Background Hepatitis B virus (HBV) infection is the major public health problem worldwide. In clinical practice, serological and molecular assays are the most commonly used diagnostic methods to detect HBV infection in clinical practices. Methods Here we present a rapid and sensitive recombinase aided amplification assay (RAA) to detect HBV at 39.0 °C for 30 min without DNA extraction from serum samples. The analytical sensitivity of RAA assay was 100 copies per reaction and showed no cross reaction with human immunodeficiency virus (HIV) and hepatitis C virus (HCV). The universality of RAA assay was validated by testing of 41 archived serum samples with predefined HBV genotypes (B, C and D). Results A total of 130 archived suspected HBV infected serum samples were detected by commercial qPCR with DNA extraction and RAA assay without DNA extraction (heat-treatment). Compared with qPCR assay as a reference, the RAA assay obtained 95.7% sensitivity and 100% specificity and a kappa value of 0.818. Conclusions We developed a rapid, convenient, highly sensitive and specific method to detect HBV without DNA extraction in clinical samples. This RAA method of HBV detection is very suitable for clinical testing.http://link.springer.com/article/10.1186/s12879-019-3814-9Hepatitis B virusDetectionRecombinase aided amplification |
| spellingShingle | Xin-xin Shen Fang-zhou Qiu Li-Ping Shen Ten-fei Yan Meng-chuan Zhao Ju-Ju Qi Chen Chen Li Zhao Le Wang Zhi-shan Feng Xue-jun Ma A rapid and sensitive recombinase aided amplification assay to detect hepatitis B virus without DNA extraction BMC Infectious Diseases Hepatitis B virus Detection Recombinase aided amplification |
| title | A rapid and sensitive recombinase aided amplification assay to detect hepatitis B virus without DNA extraction |
| title_full | A rapid and sensitive recombinase aided amplification assay to detect hepatitis B virus without DNA extraction |
| title_fullStr | A rapid and sensitive recombinase aided amplification assay to detect hepatitis B virus without DNA extraction |
| title_full_unstemmed | A rapid and sensitive recombinase aided amplification assay to detect hepatitis B virus without DNA extraction |
| title_short | A rapid and sensitive recombinase aided amplification assay to detect hepatitis B virus without DNA extraction |
| title_sort | rapid and sensitive recombinase aided amplification assay to detect hepatitis b virus without dna extraction |
| topic | Hepatitis B virus Detection Recombinase aided amplification |
| url | http://link.springer.com/article/10.1186/s12879-019-3814-9 |
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