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New isothermal amplification technique (LAMP) for direct detection of Brucella spp

Poster communication in AHVLA International Conference 2011

September 15th, 2011

Perez-Sancho M., Alvarez J., Garcia N., Martinez I., Garcia-Seco T., Gonzalez S., Goyache J. and Dominguez L.

Due to important limitations (high cost, biosafety facilities, etc.) related to bacteriological confirmation of infection by zoonotic agents such as Brucella spp., alternative tools for its direct detection in clinical samples have been developed in the last years. Among them, the LAMP (Loop-mediated isothermal amplification) technique has shown high sensitivity and specificity and can be performed without sophisticated equipment. LAMP could represent an alternative tool for Brucella spp. detection especially in developing countries where limited resources are usually available. In this context, a LAMP protocol was developed based on the IS711 target. Primers were designed using Primer ExplorerV4 software program from Eiken Chemical. Reaction mix (Bst pol, betaíne,…) and the amplification protocol was optimized as follows: first, its ability for detection of the main zoonotic Brucella species (B. suis, B. melitensis and B. abortus) was tested using bacterial DNA as template. Second, usefulness of the LAMP for direct detection of Brucella spp. was evaluated on different animal samples (lung, spleen, liver, stomach content) showing an overall sensitivity of 37.4-69.3% depending on type of sample. Our LAMP was designed based on the multiple insertion element IS711 in order to ensure a highly sensitivity of the assay, and the results show that the protocol assayed may be a useful ancillary tool for confirmation of brucellosis at a flock level, and could be carried out using only a water-bath. However, its sensitivity may be affected by type of sample so several samples should be used in order to ensure Brucella detection




Participants:

Universidad ComplutenseServicio de Zoonosis Emergentes, de Baja Prevalencia y Agresivos Biológicos (NED). Centro de Vigilancia Sanitaria Veterinaria (VISAVET). Universidad Complutense (UCM).

Universidad ComplutenseDepartamento de Sanidad Animal. Facultad de Veterinaria. Universidad Complutense (UCM).


Link to AHVLA International Conference 2011





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AHVLA International Conference 2011


AHVLA International Conference 2011
September 13rd-15th, 2011
London
United Kingdom

TITLE: New isothermal amplification technique (LAMP) for direct detection of Brucella spp


TYPE: Poster communication


AUTHORS: Perez-Sancho M., Alvarez J., Garcia N., Martinez I., Garcia-Seco T., Gonzalez S., Goyache J. and Dominguez L.


VISAVET PARTICIPANTS


First
Marta Pérez Sancho
2nd
Julio Álvarez Sánchez
3rd
Nerea García Benzaquén
4th
Irene Martínez Alares
5th
Teresa García-Seco Romero
6th
Sergio González Domínguez
7th
Joaquín Goyache Goñi
Last
Lucas Domínguez Rodríguez

DATE: September 15th, 2011


CITE THIS COMMUNICATION:

Perez-Sancho M., Alvarez J., Garcia N., Martinez I., Garcia-Seco T., Gonzalez S., Goyache J. and Dominguez L. New isothermal amplification technique (LAMP) for direct detection of Brucella spp. AHVLA International Conference 2011, Animal Health and Veterinary Laboratories Agency, London, United Kingdom, September 15th, 2011. (Poster communication)


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