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Author Martinez, J.; Simon, V.; Gonzalez, B.; Conget, P. pdf  doi
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  Title A real-time PCR-based strategy for the detection of Paenibacillus larvae vegetative cells and spores to improve the diagnosis and the screening of American foulbrood Type
  Year 2010 Publication Letters In Applied Microbiology Abbreviated Journal Lett. Appl. Microbiol.  
  Volume 50 Issue 6 Pages 603-610  
  Keywords American foulbrood; flow cytometry; Paenibacillus larvae; real-time PCR; 16S rRNA specific primers  
  Abstract Aim: To develop a real-time PCR-based strategy for the detection of Paenibacillus larvae vegetative cells and spores to improve the diagnosis and the screening of American foulbrood (AFB), the most harmful pathology of honeybee brood. Methods and Results: A real-time PCR that allowed selective identification and quantification of P. larvae 16S rRNA sequence was developed. Using standard samples quantified by flow cytometry, detection limits of 37 center dot 5 vegetative cells ml-1 and 10 spores ml-1 were determined. Compared to spread plate method, this real-time PCR-based strategy allowed, in only 2 h, the detection of P. larvae in contaminated honeys. No false-positive results were obtained. Moreover, its detection limit was 100 times lower than that of the culture method (2 vs 200 spores g-1 of honey). Conclusion: A rapid, selective, with low detection limit, sensitive and specific method to detect and quantify vegetative cells and spores of P. larvae is now available. Significance and Impact of Study: In addition to honey samples, this real-time PCR-based strategy may be also applied to confirm AFB diagnosis in honeybee brood and to screen other apiary supplies and products (bees, pollen, wax), thus broadening the control of AFB spreading.  
  Address [Martinez, J.; Simon, V.; Conget, P.] Clin Alemana Univ Desarrollo, Fac Med, Inst Ciencias, Santiago 7710162, Chile, Email: pconget@udd.cl  
  Corporate Author Thesis  
  Publisher Wiley-Blackwell Publishing, Inc Place of Publication Editor  
  Language English Summary Language Original Title  
  Series Editor Series Title Abbreviated Series Title  
  Series Volume Series Issue Edition  
  ISSN 0266-8254 ISBN Medium  
  Area Expedition Conference  
  Notes WOS:000277417000009 Approved  
  Call Number UAI @ eduardo.moreno @ Serial 89  
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