A TaqMan-based multiplex real-time PCR assay for the rapid detection of tigecycline resistance genes from bacteria, faeces and environmental samples
Abstract Background Tigecycline is a last-resort antibiotic used to treat severe infections caused by extensively drug-resistant bacteria. Recently, novel tigecycline resistance genes tet(X3) and tet(X4) have been reported, which pose a great challenge to human health and food security. The current...
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doaj-91ed6052e16841d5ab02626de71ecc102020-11-25T03:52:53ZengBMCBMC Microbiology1471-21802020-06-012011710.1186/s12866-020-01813-8A TaqMan-based multiplex real-time PCR assay for the rapid detection of tigecycline resistance genes from bacteria, faeces and environmental samplesYiming Li0Zhangqi Shen1Shuangyang Ding2Shaolin Wang3Beijing Advanced Innovation Center for Food Nutrition and Human Health, College of Veterinary Medicine, China Agricultural UniversityBeijing Advanced Innovation Center for Food Nutrition and Human Health, College of Veterinary Medicine, China Agricultural UniversityBeijing Advanced Innovation Center for Food Nutrition and Human Health, College of Veterinary Medicine, China Agricultural UniversityBeijing Advanced Innovation Center for Food Nutrition and Human Health, College of Veterinary Medicine, China Agricultural UniversityAbstract Background Tigecycline is a last-resort antibiotic used to treat severe infections caused by extensively drug-resistant bacteria. Recently, novel tigecycline resistance genes tet(X3) and tet(X4) have been reported, which pose a great challenge to human health and food security. The current study aimed to establish a TaqMan-based real-time PCR assay for the rapid detection of the tigecycline-resistant genes tet(X3) and tet(X4). Results No false-positive result was found, and the results of the TaqMan-based real-time PCR assay showed 100% concordance with the results of the sequencing analyses. This proposed method can detect the two genes at the level of 1 × 102 copies/μL, and the whole process is completed within an hour, allowing rapid screening of tet(X3) and tet(X4) genes in cultured bacteria, faeces, and soil samples. Conclusion Taken together, the TaqMan-based real-time PCR method established in this study is rapid, sensitive, specific, and is capable of detecting the two genes not only in bacteria, but also in environmental samples.http://link.springer.com/article/10.1186/s12866-020-01813-8Tigecycline resistanceTaqManReal-time PCR |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Yiming Li Zhangqi Shen Shuangyang Ding Shaolin Wang |
spellingShingle |
Yiming Li Zhangqi Shen Shuangyang Ding Shaolin Wang A TaqMan-based multiplex real-time PCR assay for the rapid detection of tigecycline resistance genes from bacteria, faeces and environmental samples BMC Microbiology Tigecycline resistance TaqMan Real-time PCR |
author_facet |
Yiming Li Zhangqi Shen Shuangyang Ding Shaolin Wang |
author_sort |
Yiming Li |
title |
A TaqMan-based multiplex real-time PCR assay for the rapid detection of tigecycline resistance genes from bacteria, faeces and environmental samples |
title_short |
A TaqMan-based multiplex real-time PCR assay for the rapid detection of tigecycline resistance genes from bacteria, faeces and environmental samples |
title_full |
A TaqMan-based multiplex real-time PCR assay for the rapid detection of tigecycline resistance genes from bacteria, faeces and environmental samples |
title_fullStr |
A TaqMan-based multiplex real-time PCR assay for the rapid detection of tigecycline resistance genes from bacteria, faeces and environmental samples |
title_full_unstemmed |
A TaqMan-based multiplex real-time PCR assay for the rapid detection of tigecycline resistance genes from bacteria, faeces and environmental samples |
title_sort |
taqman-based multiplex real-time pcr assay for the rapid detection of tigecycline resistance genes from bacteria, faeces and environmental samples |
publisher |
BMC |
series |
BMC Microbiology |
issn |
1471-2180 |
publishDate |
2020-06-01 |
description |
Abstract Background Tigecycline is a last-resort antibiotic used to treat severe infections caused by extensively drug-resistant bacteria. Recently, novel tigecycline resistance genes tet(X3) and tet(X4) have been reported, which pose a great challenge to human health and food security. The current study aimed to establish a TaqMan-based real-time PCR assay for the rapid detection of the tigecycline-resistant genes tet(X3) and tet(X4). Results No false-positive result was found, and the results of the TaqMan-based real-time PCR assay showed 100% concordance with the results of the sequencing analyses. This proposed method can detect the two genes at the level of 1 × 102 copies/μL, and the whole process is completed within an hour, allowing rapid screening of tet(X3) and tet(X4) genes in cultured bacteria, faeces, and soil samples. Conclusion Taken together, the TaqMan-based real-time PCR method established in this study is rapid, sensitive, specific, and is capable of detecting the two genes not only in bacteria, but also in environmental samples. |
topic |
Tigecycline resistance TaqMan Real-time PCR |
url |
http://link.springer.com/article/10.1186/s12866-020-01813-8 |
work_keys_str_mv |
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