TY - JOUR
T1 - Development and validation of an insulated isothermal PCR assay for the rapid detection of Mannheimia haemolytica
AU - Dao, Xiaofang
AU - Hung, Chien Che
AU - Yang, Youwen
AU - Wang, Juan
AU - Yang, Falong
N1 - Publisher Copyright:
© 2022 The Author(s).
PY - 2022/3
Y1 - 2022/3
N2 - We developed a rapid insulated isothermal PCR (iiPCR) assay for on-site detection of Mannheimia haemolytica using a primer and probe set targeting the superoxide dismutase (sodA) gene. Our iiPCR assay detected M. haemolytica clinical isolates successfully and produced negative results on other bovine or ovine respiratory pathogens, including Histophilus somni, Bibersteinia trehalosi, Trueperella pyogenes, Streptococcus suis, and Mycoplasma spp., indicating that the PCR reactions were specific. Additionally, our iiPCR assay detected as few as 21 copies of genomic DNA and 17.2 cfu/mL of bacterial culture, which was 10 and 100 times more sensitive than conventional PCR, respectively. Our iiPCR assay can be performed on a portable device in a total of 58 min and may be a useful tool for the detection of M. haemolytica in bovine and ovine respiratory disease in the field.
AB - We developed a rapid insulated isothermal PCR (iiPCR) assay for on-site detection of Mannheimia haemolytica using a primer and probe set targeting the superoxide dismutase (sodA) gene. Our iiPCR assay detected M. haemolytica clinical isolates successfully and produced negative results on other bovine or ovine respiratory pathogens, including Histophilus somni, Bibersteinia trehalosi, Trueperella pyogenes, Streptococcus suis, and Mycoplasma spp., indicating that the PCR reactions were specific. Additionally, our iiPCR assay detected as few as 21 copies of genomic DNA and 17.2 cfu/mL of bacterial culture, which was 10 and 100 times more sensitive than conventional PCR, respectively. Our iiPCR assay can be performed on a portable device in a total of 58 min and may be a useful tool for the detection of M. haemolytica in bovine and ovine respiratory disease in the field.
KW - Mannheimia haemolytica
KW - bovine respiratory disease
KW - insulated isothermal PCR
KW - on-site detection
UR - http://www.scopus.com/inward/record.url?scp=85125074650&partnerID=8YFLogxK
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U2 - 10.1177/10406387211068447
DO - 10.1177/10406387211068447
M3 - Article
C2 - 35139720
AN - SCOPUS:85125074650
SN - 1040-6387
VL - 34
SP - 302
EP - 305
JO - Journal of Veterinary Diagnostic Investigation
JF - Journal of Veterinary Diagnostic Investigation
IS - 2
ER -