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A sensitive one-step real-time RT-PCR method for detection of deformed wing virus and black queen cell virus in honeybee Apis mellifera

Journal of Virological Methods publish this investigation article

February 1st, 2008

A one-step real-time RT-PCR based on SYBR Green (SG) chemistry was developed for the detection, differentiation and quantification of two of the most common viruses on the honeybee Apis mellifera L., deformed wing virus and black queen cell virus. Two sets of primers specific for each virus, were designed in conserved regions of the viral genome for their use in the one-step real-time RT-PCR. Both reactions were optimized for highest sensitivity and specificity and SG-based real-time was used to achieve quantitative detection. All samples evaluated in this study were from Spanish honeybee colonies. Viral detection and identification was confirmed by sequencing of the PCR products. The described one-step real-time SG RT-PCR proved to be a fast, accurate and useful technique to detect and even quantify these honeybee viruses that cause unapparent infections, and might contribute with other factors to the increasing honeybee colonies depopulation




Kukielka D., Esperon F., Higes M. and Sanchez-Vizcaino JM..




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A sensitive one-step real-time RT-PCR method for detection of deformed wing virus and black queen cell virus in honeybee Apis mellifera

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A sensitive one-step real-time RT-PCR method for detection of deformed wing virus and black queen cell virus in honeybee Apis mellifera



Participants:

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

Universidad ComplutenseServicio de Inmunología Viral y Medicina Preventiva (SUAT). Centro de Vigilancia Sanitaria Veterinaria (VISAVET). Universidad Complutense (UCM).

Gobierno de Castilla-La ManchaCentro Apícola Regional (CAR). Gobierno de Castilla-La Mancha (JCCM).







Journal of Virological Methods
FACTOR YEAR Q
2.077 2008

NLMID: 8005839

PMID: 17964669

ISSN: 0166-0934



TITLE: A sensitive one-step real-time RT-PCR method for detection of deformed wing virus and black queen cell virus in honeybee Apis mellifera


JOURNAL: J. Virol. Methods


NUMERACIÓN: 147(2):275-81


AÑO: 2008


PUBLISHER: Elsevier/North-Holland Biomedical Press


AUTHORS: Kukielka D., Esperon F., Higes M. and Sanchez-Vizcaino JM..


VISAVET PARTICIPANTS


José Manuel Sánchez-Vizcaíno Rodríguez

DOI: https://doi.org/10.1016/j.jviromet.2007.09.008


CITE THIS PUBLICATION:

Kukielka D., Esperon F., Higes M. and Sanchez-Vizcaino JM. A sensitive one-step real-time RT-PCR method for detection of deformed wing virus and black queen cell virus in honeybee Apis mellifera. Journal of Virological Methods. 147(2):275-81. 2008. (A). ISSN: 0166-0934. DOI: 10.1016/j.jviromet.2007.09.008


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