| Autor: |
Rui-qing Zhang, Zheng Li, Gui-xia Li, Yan-qing Tie, Xin-na Li, Yuan Gao, Qing-xia Duan, Le Wang, Li Zhao, Guo-hao Fan, Xue-ding Bai, Rui-huan Wang, Zi-wei Chen, Jin-rong Wang, Yong Wu, Meng-chuan Zhao, Zhi-shan Feng, Ji Wang, Xue-jun Ma |
| Jazyk: |
angličtina |
| Rok vydání: |
2020 |
| Předmět: |
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| Zdroj: |
International Journal of Infectious Diseases, Vol 93, Iss , Pp 224-230 (2020) |
| Druh dokumentu: |
article |
| ISSN: |
1201-9712 |
| DOI: |
10.1016/j.ijid.2020.01.053 |
| Popis: |
Objectives: Bordetella pertussis is a highly contagious respiratory agent and is the causative pathogen of pertussis, which primarily affects children. Current diagnostic techniques for this pathogen have a variety of limitations including a long culture time, low bacterial load, and lack of specificity. Methods: This article reports the development of a one-tube nested quantitative real-time PCR assay using the locked nucleic acid (LNA) technique (LNA-OTN-q-PCR), targeting the BP485 gene and using a simple inexpensive extraction method. A total of 130 clinical samples from patients with clinically suspected pertussis, collected from the Children’s Hospital of Hebei, China, were tested by LNA-OTN-q-PCR assay. RT-PCR and two-step semi-nested PCR assays were performed in parallel for comparison. Results: Only strains of B. pertussis were identified as positive, whereas all of the remaining strains were appropriately identified as negative by the LNA-OTN-q-PCR assay. A single copy per reaction can be detected by the LNA-OTN-q-PCR assay. Additionally, the sensitivity of this method was 100 times that of the RT-PCR assay (100 copies per reaction). Sixty-three of the 130 clinical samples were detected positive by LNA-OTN-q-PCR assay; in contrast, RT-PCR was able to detect only 41 positive samples. Following this, all 63 samples were positively identified by two-step semi-nested PCR. Compared with the two-step semi-nested PCR assay, both the specificity and sensitivity of the LNA-OTN-q-PCR assay using purified DNA and crude extract were 100%. Conclusions: This assay was able to detect B. pertussis infection with high sensitivity and specificity. This test shows great potential as a promising technique to detect B. pertussis in both clinical laboratories and public health settings. Keywords: One-tube nested quantitative real-time PCR using the LNA technique, Bordetella pertussis, Simple extraction method |
| Databáze: |
Directory of Open Access Journals |
| Externí odkaz: |
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