title: Loop-mediated isothermal amplification for detection of Staphylococcus aureus in dairy cow suffering from mastitis.
abstract: To develop a rapid detection method of Staphylococcus aureus using loop-mediated isothermal amplification (LAMP), four specific primers were designed according to six distinct sequences of the nuc gene. In addition, the specificity and sensitivity of LAMP were verified and compared with those of PCR. Results showed that the LAMP reaction was completed within 45 min at 62.5°C, and ladder bands were appeared in LAMP products analyzed by gel electrophoresis. After adding 1x SYBR Green l, the positive reaction tube showed green color and the negative reaction tube remained orange, indicating that the LAMP has high specificity. The minimal detectable concentration of LAMP was 1 × 10² CFU/mL and that of PCR was 1 × 10⁴ CFU/mL, indicating that the LAMP was 100 times more sensitive than the PCR. The LAMP method for detection of Staphylococcus aureus has many advantages, such as simple operation, high sensitivity, high specificity, and rapid analysis. Therefore, this method is more suitable for the rapid on-site detection of Staphylococcus aureus.
authors.affiliation: The College of Traditional Chinese Veterinary Medicine, Agricultural University of Hebei, Dingzhou 073000, China.
authors.forename: Zhang
authors.initials: Z
authors.lastname: Tie
authors.forename: Wang
authors.initials: W
authors.lastname: Chunguang
authors.forename: Wei
authors.initials: W
authors.lastname: Xiaoyuan
authors.forename: Zhao
authors.initials: Z
authors.lastname: Xinghua
authors.forename: Zhong
authors.initials: Z
authors.lastname: Xiuhui
mesh_terms: D000818:Animals; D052160:Benzothiazoles; D002417:Cattle; D003612:Dairying; D003959:Diamines; D005260:Female; D008413:Mastitis / Q000382:microbiology*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D009930:Organic Chemicals / Q000378:metabolism; D011804:Quinolines; D012680:Sensitivity and Specificity; D013211:Staphylococcus aureus / Q000235:genetics* / Q000302:isolation & purification*
chemical_list: D052160:Benzothiazoles; D003959:Diamines; D009930:Organic Chemicals; D011804:Quinolines; C098022:SYBR Green I