TY - JOUR
T1 - A novel approach for detection of brucella using a real-time recombinase polymerase amplification assay
AU - Qin, Lide
AU - Nan, Wenlong
AU - Wang, Yong
AU - Zhang, Yueyong
AU - Tan, Pengfei
AU - Chen, Yuqi
AU - Mao, Kairong
AU - Chen, Yiping
N1 - Funding Information:
This work was financially supported by the National Key Research and Development Program of China (No. 2017YFF0208600 ).
Publisher Copyright:
© 2019
PY - 2019/12
Y1 - 2019/12
N2 - Brucella, the etiological agent of brucellosis, is an important zoonosis pathogen worldwide. Brucella infects humans and various domestic and wild animals, and represents a great threat to public health and animal husbandry. In the present study, we developed a real-time recombinase polymerase amplification (RPA) assay for the detection of Brucella. The assay targeted the bcsp31 gene of Brucella, and an RPA exo probe and a pair of primers were selected for assay validation. RPA sensitivity and specificity were evaluated using plasmid standards, Brucella representative strains, and non-Brucella strains. The RPA assay achieved a detection limit of 17 molecules in 95% of cases based on probit analysis, and could successfully distinguish 18 representative Brucella strains (B. abortus biovars 1, 2, 3, 4, 5, 6, 7 and 9, B. melitensis biovars 1, 2 and 3, B. suis biovars 1, 2, 3 and 4, B. canis, B. neotomae and B. ovis), and four Brucella vaccine strains (A19, S19, S2 and M5). A total of 52 Brucella field strains were detected by real-time PCR and RPA in parallel, and compared with real-time PCR, the sensitivity of the RPA assay was 94% (49/52). Thus, this RPA assay may be a rapid, sensitive, and specific tool for the prevention and control of Brucellosis.
AB - Brucella, the etiological agent of brucellosis, is an important zoonosis pathogen worldwide. Brucella infects humans and various domestic and wild animals, and represents a great threat to public health and animal husbandry. In the present study, we developed a real-time recombinase polymerase amplification (RPA) assay for the detection of Brucella. The assay targeted the bcsp31 gene of Brucella, and an RPA exo probe and a pair of primers were selected for assay validation. RPA sensitivity and specificity were evaluated using plasmid standards, Brucella representative strains, and non-Brucella strains. The RPA assay achieved a detection limit of 17 molecules in 95% of cases based on probit analysis, and could successfully distinguish 18 representative Brucella strains (B. abortus biovars 1, 2, 3, 4, 5, 6, 7 and 9, B. melitensis biovars 1, 2 and 3, B. suis biovars 1, 2, 3 and 4, B. canis, B. neotomae and B. ovis), and four Brucella vaccine strains (A19, S19, S2 and M5). A total of 52 Brucella field strains were detected by real-time PCR and RPA in parallel, and compared with real-time PCR, the sensitivity of the RPA assay was 94% (49/52). Thus, this RPA assay may be a rapid, sensitive, and specific tool for the prevention and control of Brucellosis.
KW - bcsp31 gene
KW - Brucella
KW - Exo probe
KW - RPA
UR - http://www.scopus.com/inward/record.url?scp=85072997557&partnerID=8YFLogxK
U2 - 10.1016/j.mcp.2019.101451
DO - 10.1016/j.mcp.2019.101451
M3 - Article
C2 - 31541671
AN - SCOPUS:85072997557
SN - 0890-8508
VL - 48
JO - Molecular and Cellular Probes
JF - Molecular and Cellular Probes
M1 - 101451
ER -
