Unknown

Dataset Information

0

Rapid visual detection of Mycobacterium avium subsp. paratuberculosis by recombinase polymerase amplification combined with a lateral flow dipstick.


ABSTRACT: Paratuberculosis (Johne's disease) is a chronic debilitating disease of domestic and wild ruminants. However, widespread point-of-care testing is infrequent due to the lack of a robust method. The isothermal recombinase polymerase amplification (RPA) technique has applied for rapid diagnosis. Herein, RPA combined with a lateral flow dipstick (LFD) assay was developed to estimate DNA from Mycobacterium avium subsp. paratuberculosis. First, analytical specificity and sensitivity of the RPA-nfo primer and probe sets were assessed. The assay successfully detected M. paratuberculosis DNA in 30 min at 39°C with a detection limit of up to eight copies per reaction, which was equivalent to that of the real-time quantitative polymerase chain reaction (qPCR) assay. The assay was specific, as it did not amplify genomes from five other Mycobacterium spp. or five pathogenic enteric bacteria. Six hundred-twelve clinical samples (320 fecal and 292 serum) were assessed by RPA-LFD, qPCR, and enzyme-linked immunosorbent assay, respectively. The RPA-LFD assay yielded 100% sensitivity, 97.63% specificity, and 98.44% concordance rate with the qPCR results. This is the first report utilizing an RPA-LFD assay to visualize and rapidly detect M. paratuberculosis. Our results show this assay should be a useful method for the diagnosis of paratuberculosis in resource-constrained settings.

SUBMITTER: Zhao G 

PROVIDER: S-EPMC5879072 | biostudies-literature | 2018 Mar

REPOSITORIES: biostudies-literature

altmetric image

Publications

Rapid visual detection of <i>Mycobacterium avium</i> subsp. <i>paratuberculosis</i> by recombinase polymerase amplification combined with a lateral flow dipstick.

Zhao Guimin G   Wang Hongmei H   Hou Peili P   He Chengqiang C   He Hongbin H  

Journal of veterinary science 20180301 2


Paratuberculosis (Johne's disease) is a chronic debilitating disease of domestic and wild ruminants. However, widespread point-of-care testing is infrequent due to the lack of a robust method. The isothermal recombinase polymerase amplification (RPA) technique has applied for rapid diagnosis. Herein, RPA combined with a lateral flow dipstick (LFD) assay was developed to estimate DNA from <i>Mycobacterium avium</i> subsp. <i>paratuberculosis</i>. First, analytical specificity and sensitivity of t  ...[more]

Similar Datasets

| S-EPMC5167419 | biostudies-literature
| S-EPMC5006264 | biostudies-literature
| S-EPMC7203717 | biostudies-literature
| S-EPMC10197247 | biostudies-literature
| S-EPMC4810606 | biostudies-literature
| S-EPMC88986 | biostudies-literature
| S-EPMC156888 | biostudies-literature
| S-EPMC140397 | biostudies-literature
| S-EPMC9263184 | biostudies-literature
| S-EPMC6429820 | biostudies-literature