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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.
Lett Appl Microbiol. 2010 Jun 01; 50(6):603-10.LA

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 37.5 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.

Authors+Show Affiliations

Instituto de Ciencias, Facultad de Medicina, Clinica Alemana Universidad del Desarrollo, Santiago, Chile.No affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Evaluation Study
Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

20406378

Citation

Martínez, J, et al. "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." Letters in Applied Microbiology, vol. 50, no. 6, 2010, pp. 603-10.
Martínez J, Simon V, Gonzalez B, et al. 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. Lett Appl Microbiol. 2010;50(6):603-10.
Martínez, J., Simon, V., Gonzalez, B., & Conget, P. (2010). 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. Letters in Applied Microbiology, 50(6), 603-10. https://doi.org/10.1111/j.1472-765X.2010.02840.x
Martínez J, et al. 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. Lett Appl Microbiol. 2010 Jun 1;50(6):603-10. PubMed PMID: 20406378.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - 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. AU - Martínez,J, AU - Simon,V, AU - Gonzalez,B, AU - Conget,P, Y1 - 2010/03/19/ PY - 2010/4/22/entrez PY - 2010/4/22/pubmed PY - 2010/9/21/medline SP - 603 EP - 10 JF - Letters in applied microbiology JO - Lett. Appl. Microbiol. VL - 50 IS - 6 N2 - 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 37.5 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. SN - 1472-765X UR - https://www.unboundmedicine.com/medline/citation/20406378/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_ L2 - https://doi.org/10.1111/j.1472-765X.2010.02840.x DB - PRIME DP - Unbound Medicine ER -