Ultra-rapid real-time PCR for the detection of Paenibacillus larvae, the causative agent of American Foulbrood (AFB).
A novel micro-PCR-based detection method, termed ultra-rapid real-time PCR, was applied to the development of a rapid detection for Paenibacillus larvae (P. larvae) which is the causative agent of American Foulbrood (AFB). This method was designed to detect the 16S rRNA gene of P. larvae with a micro-scale chip-based real-time PCR system, GenSpector TMC-1000, which has uncommonly fast heating and cooling rates (10 degrees C per second) and small reaction volume (6microl). In the application of ultra-rapid real-time PCR detection to an AFB-infected larva, the minimum detection time was 7 min and 54s total reaction time (30 cycles), including the melting temperature analysis. To the best of our knowledge, this novel detection method is one of the most rapid real-time PCR-based detection tools.
Han
Sang-Hoon
SH
Department of Life Science, College of Natural Science, Kyonggi University, Suwon, Kyonggi-do, Republic of Korea.
Lee
Do-Bu
DB
Lee
Dong-Woo
DW
Kim
Eul-Hwan
EH
Yoon
Byoung-Su
BS
D000818:Animals; D001406:Bacillaceae / Q000235:genetics / Q000302:isolation & purification*; D001516:Bees / Q000382:microbiology*; D004269:DNA, Bacterial / Q000032:analysis; D016908:Gram-Positive Bacterial Infections / Q000382:microbiology / Q000662:veterinary*; D054884:Host-Pathogen Interactions; D007814:Larva / Q000382:microbiology; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013997:Time Factors
D004269:DNA, Bacterial