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| Loop-mediated Isothermal Amplification for Sensitive and Rapid Detection of Mycoplasma hyopneumoniae |
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Abstract Abstract Using the loop-mediated isothermal amplification(LAMP), a novel method of gene amplification for Mycoplasma hyopneumoniae detection was established. A set of six primers was designed specificity to recognize the 16S rRNA gene, which is a relatively conserved region of M. hyopneumoniae. The LAMP-based assay could be completed within 30 min at 63℃. Results showed that the LAMP-based assay, compared with PCR, had high superiority. The detection limit of the LAMP assay was found to be 10 copies per reaction determined by using a recombined plasmid containing the target sequence, which was 10-fold higher than that of the PCR assay. In addition, both LAMP and PCR were highly specific to M. hyopneumoniae. Furthermore, the LAMP and PCR assay was applied to 127 clinical nasal swab samples collected from pig farms. For the LAMP assay, all the nasal swab specimens tested positive, while for the PCR assay, 116 nasal swab specimens tested positive. The data confirmed that LAMP was more sensitive than PCR for the detection of clinical samples. The LAMP assay is easy to perform, extremely rapid, highly sensitive, specificity and cost-effective, therefore it has the great potentiality suitable for diagnosis of M. hyopneumoniae both in well-equipped laboratories and in field situations.
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Received: 03 April 2009
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NI He-Jia, CHENG Xue, WANG Jing, GAO Ze-Ping, WEI Pan-Pan, TAN Jia-Li, SUN Jing, LI Hai-Tao, GAO Ji-Guo. Isolation and Screening of High Virulent Bacillus thuringiensis Strains Against Lepidopteran Pests in Some Areas of Liaoning Province[J]. 农业生物技术学报, 2019, 27(8): 1513-1520. |
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