Development of a novel LAMP diagnostic method for visible detection of swine Pasteurella multocida.
A set of four specific primers for six regions of kmt1 gene from a species specific region was designed for developing the loop-mediated isothermal amplification diagnostic method of swine Pasteurella multocida (Pm-LAMP). After the Pm-LAMP was carried out at 63°C for 1 h, the LAMP products could be visually confirmed using fluorescent dyes as detection reagent under UV-illumination. In sensitivity, the detection limit of the Pm-LAMP was 10 cfu/mL, and was 1 log less than that of the PCR method. In specificity, the Pm-LAMP did not amplify genomic DNA of swine common respiratory pathogens. Furthermore, based on results for clinical swab samples (n = 31) using PCR detection as golden standard, relative sensitivity of the Pm-LAMP was 100%, relative specificity of the Pm-LAMP was 90.9%, and percentage of observation agreement was 93.5% (Kappa = 0.85). The Pm-LAMP method should be a useful diagnostic tool for rapid and visible detection of swine Pasteurella multocida.
Sun
Dongbo
D
College of Animal Science and Veterinary Medicine, Heilongjiang Bayi Agricultural University, Daqing High-tech Industrial Development Zone, Daqing, 163319, People's Republic of China. dongbosun@yahoo.com.cn
Wang
Jianfa
J
Wu
Rui
R
Wang
Chunren
C
He
Xianjing
X
Zheng
Jiasan
J
Yang
Huanmin
H
D000818:Animals; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D021141:Nucleic Acid Amplification Techniques / Q000379:methods / Q000662:veterinary*; D010326:Pasteurella Infections / Q000175:diagnosis / Q000382:microbiology / Q000662:veterinary*; D016979:Pasteurella multocida / Q000235:genetics / Q000302:isolation & purification*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013552:Swine; D013553:Swine Diseases / Q000175:diagnosis / Q000382:microbiology*
D004269:DNA, Bacterial