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| Content Provider | Springer Nature : BioMed Central |
|---|---|
| Author | Thanh, Tran Tan Anh, Nguyen To Tham, Nguyen Thi Van, Hoang Minh Tu Sabanathan, Saraswathy Qui, Phan Tu Ngan, Tran Thuy Van, Tran Thi My Nguyet, Lam Anh Ny, Nguyen Thi Han Thanh, Le Thi My Chai, Ong Kien Perera, David Viet, Do Chau Khanh, Truong Huu Ha, Do Quang Tuan, Ha Manh Wong, Kum Thong Hung, Nguyen Thanh Chau, Nguyen Van Vinh Thwaites, Guy van Doorn, H Rogier Van Tan, Le |
| Abstract | Background Hand foot and mouth disease (HFMD) is a disease of public health importance across the Asia-Pacific region. The disease is caused by enteroviruses (EVs), in particular enterovirus A71 (EV-A71). In EV-A71-associated HFMD, the infection is sometimes associated with severe manifestations including neurological involvement and fatal outcome. The availability of a robust diagnostic assay to distinguish EV-A71 from other EVs is important for patient management and outbreak response. Methods We developed and validated an internally controlled one-step single-tube real-time RT-PCR in terms of sensitivity, linearity, precision, and specificity for simultaneous detection of EVs and EV-A71. Subsequently, the assay was then applied on throat and rectal swabs sampled from 434 HFMD patients. Results The assay was evaluated using both plasmid DNA and viral RNA and has shown to be reproducible with a maximum assay variation of 4.41 % and sensitive with a limit of detection less than 10 copies of target template per reaction, while cross-reactivity with other EV serotypes was not observed. When compared against a published VP1 nested RT-PCR using 112 diagnostic throat and rectal swabs from 112 children with a clinical diagnosis of HFMD during 2014, the multiplex assay had a higher sensitivity and 100 % concordance with sequencing results which showed EVs in 77/112 (68.8 %) and EV-A71 in 7/112 (6.3 %). When applied to clinical diagnostics for 322 children, the assay detected EVs in throat swabs of 257/322 (79.8 %) of which EV-A71 was detected in 36/322 (11.2 %) children. The detection rate increased to 93.5 % (301/322) and 13.4 % (43/322) for EVs and EV-A71, respectively, when rectal swabs from 65 throat-negative children were further analyzed. Conclusion We have successfully developed and validated a sensitive internally controlled multiplex assay for rapid detection of EVs and EV-A71, which is useful for clinical management and outbreak control of HFMD. |
| Related Links | https://virologyj.biomedcentral.com/counter/pdf/10.1186/s12985-015-0316-2.pdf |
| Ending Page | 9 |
| Page Count | 9 |
| Starting Page | 1 |
| File Format | HTM / HTML |
| DOI | 10.1186/s12985-015-0316-2 |
| Journal | Virology Journal |
| Issue Number | 1 |
| Volume Number | 12 |
| Language | English |
| Publisher | BioMed Central |
| Publisher Date | 2015-06-09 |
| Access Restriction | Open |
| Subject Keyword | Virology Hand foot and mouth disease Enteroviruses Enterovirus A71 Real-time RT-PCR Diagnosis |
| Content Type | Text |
| Resource Type | Article |
| Subject | Virology Infectious Diseases |
| Journal Impact Factor | 4/2023 |
| 5-Year Journal Impact Factor | 3.8/2023 |
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