DNA-Based Detection and Identification of Eight Mastitis Pathogens

摘要

LAMP primer sets for detecting eight mastitis pathogens are disclosed. Methods and kits of using the primer sets to simultaneously detecting at least two of the eight mastitis pathogens are also described.

主权项

1. A nucleotide primer set for LAMP amplification, used for detecting E. coli comprising a set selected from the group consisting of:
(i) a nucleotide primer set for detecting E. coli comprising
(a) a F3 primer having a sequence identical or substantially identical to SEQ ID NO:1,(b) a B3 primer having a sequence identical or substantially identical to SEQ ID NO:2,(c) a FIP primer having a sequence identical or substantially identical to SEQ ID NO:3, and(d) a BIP primer having a sequence identical or substantially identical to SEQ ID NO:4, and optionally(e) a LF primer having a sequence identical or substantially identical to SEQ ID NO:5, and(f) a LB primer having a sequence identical or substantially identical to SEQ ID NO:6; (ii) a nucleotide primer set for LAMP amplification, used for detecting Staphylococcus aureus comprising
(a) a F3 primer having a sequence identical or substantially identical to SEQ ID NO:7,(b) a B3 primer having a sequence identical or substantially identical to SEQ ID NO:8,(c) a FIP primer having a sequence identical or substantially identical to SEQ ID NO:9, and(d) a BIP primer having a sequence identical or substantially identical to SEQ ID NO:10; (iii) a nucleotide primer set for LAMP amplification, used for detecting Klebsiella pneumonia comprising
(a) a F3 primer having a sequence identical or substantially identical to SEQ ID NO:11,(b) a B3 primer having a sequence identical or substantially identical to SEQ ID NO:12,(c) a FIP primer having a sequence identical or substantially identical to SEQ ID NO:13, and(d) a BIP primer having a sequence identical or substantially identical to SEQ ID NO:14, and optionally(e) a LF primer having a sequence identical or substantially identical to SEQ ID NO:15, and(f) a LB primer having a sequence identical or substantially identical to SEQ ID NO:16; (iv) a nucleotide primer set for LAMP amplification, used for detecting Streptococcus uberis comprising
(a) a F3 primer having a sequence identical or substantially identical to SEQ ID NO:17,(b) a B3 primer having a sequence identical or substantially identical to SEQ ID NO:18,(c) a FIP primer having a sequence identical or substantially identical to SEQ ID NO:19, and(d) a BIP primer having a sequence identical or substantially identical to SEQ ID NO:20, and optionally(e) a LF primer having a sequence identical or substantially identical to SEQ ID NO:21, and(f) a LB primer having a sequence identical or substantially identical to SEQ ID NO:22; (v) a nucleotide primer set for LAMP amplification, used for detecting Streptococcus dysgalactiae comprising
(a) a F3 primer having a sequence identical or substantially identical to SEQ ID NO:23,(b) a B3 primer having a sequence identical or substantially identical to SEQ ID NO:24,(c) a FIP primer having a sequence identical or substantially identical to SEQ ID NO:25, and(d) a BIP primer having a sequence identical or substantially identical to SEQ ID NO:26, and optionally(e) a LF primer having a sequence identical or substantially identical to SEQ ID NO:27, and(f) a LB primer having a sequence identical or substantially identical to SEQ ID NO:28; (vi) a nucleotide primer set for LAMP amplification, used for detecting Streptococcus agalactiae comprising
(a) a F3 primer having a sequence identical or substantially identical to SEQ ID NO:29,(b) a B3 primer having a sequence identical or substantially identical to SEQ ID NO:30,(c) a FIP primer having a sequence identical or substantially identical to SEQ ID NO:31, and(d) a BIP primer having a sequence identical or substantially identical to SEQ ID NO:32; (vii) a nucleotide primer set for LAMP amplification, used for detecting Mycoplasma bovis comprising
(a) a F3 primer having a sequence identical or substantially identical to SEQ ID NO:33,(b) a B3 primer having a sequence identical or substantially identical to SEQ ID NO:34,(c) a FIP primer having a sequence identical or substantially identical to SEQ ID NO:35, and(d) a BIP primer having a sequence identical or substantially identical to SEQ ID NO:36, and optionally(e) a LF primer having a sequence identical or substantially identical to SEQ ID NO:37, and(f) a LB primer having a sequence identical or substantially identical to SEQ ID NO:38; and (viii) a nucleotide primer set for LAMP amplification, used for detecting coagulase-negative Staphylococci (CNS) tuf or any one of the subtypes including S. epidermidis, S. chromogenes, and S. simulans, comprising
(a) a F3 primer having a sequence identical or substantially identical to SEQ ID NO:39,(b) a B3 primer having a sequence identical or substantially identical to SEQ ID NO:40,(c) a FIP primer having a sequence identical or substantially identical to SEQ ID NO:41, and(d) a BIP primer having a sequence identical or substantially identical to SEQ ID NO:42.