PMID- 19559729
OWN - NLM
STAT- MEDLINE
DCOM- 20091109
LR  - 20090828
IS  - 1879-0984 (Electronic)
IS  - 0166-0934 (Linking)
VI  - 161
IP  - 2
DP  - 2009 Nov
TI  - One-step real-time quantitative PCR assays for the detection and field study of 
      Sacbrood honeybee and Acute bee paralysis viruses.
PG  - 240-6
LID - 10.1016/j.jviromet.2009.06.014 [doi]
AB  - Two one-step real-time RT-PCR assays, based on SYBR Green (SG) chemistry, were 
      developed or adapted respectively, for the detection, differentiation, and 
      quantitation of two important honeybee viruses: Sacbrood virus (SBV) and Acute 
      bee paralysis virus (ABPV). Both reactions were optimized to yield the highest 
      sensitivity and specificity. The genome equivalent copies (GEC) detection limit 
      per reaction was 389.3 for the ABPV RT-PCR. The GEC detection limit per reaction 
      was 298.9 for the SBV RT-PCR. Viral detection and identification were confirmed 
      by melting curve analysis and sequencing of the PCR products. Both techniques 
      were used to evaluate Spanish field samples and establish the distribution of 
      these viruses. Acute bee paralysis virus was not detected, and Sacbrood virus was 
      present at low frequencies. The one-step real-time SG RT-PCR methods are fast, 
      accurate, and useful for detecting and quantifying these honeybee viruses, which 
      cause inapparent infections and contribute to the increasing depopulation of 
      honeybee colonies.
FAU - Kukielka, Deborah
AU  - Kukielka D
AD  - Departamento de Sanidad Animal Facultad de Veterinaria, UCM, Avda Puerta de 
      Hierro s/n, Madrid, Spain. deborah@sanidadanimal.info
FAU - Sanchez-Vizcaino, Jose Manuel
AU  - Sanchez-Vizcaino JM
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20090625
PL  - Netherlands
TA  - J Virol Methods
JT  - Journal of virological methods
JID - 8005839
RN  - 0 (RNA, Viral)
SB  - IM
MH  - Animals
MH  - Bees/*virology
MH  - *Insect Viruses/genetics/isolation & purification
MH  - Phylogeny
MH  - RNA, Viral/analysis/genetics
MH  - Reverse Transcriptase Polymerase Chain Reaction/methods
MH  - Sensitivity and Specificity
MH  - Sequence Alignment
MH  - Sequence Analysis, DNA
MH  - Spain
EDAT- 2009/06/30 09:00
MHDA- 2009/11/10 06:00
CRDT- 2009/06/30 09:00
PHST- 2009/03/16 00:00 [received]
PHST- 2009/06/12 00:00 [revised]
PHST- 2009/06/16 00:00 [accepted]
PHST- 2009/06/30 09:00 [entrez]
PHST- 2009/06/30 09:00 [pubmed]
PHST- 2009/11/10 06:00 [medline]
AID - S0166-0934(09)00303-6 [pii]
AID - 10.1016/j.jviromet.2009.06.014 [doi]
PST - ppublish
SO  - J Virol Methods. 2009 Nov;161(2):240-6. doi: 10.1016/j.jviromet.2009.06.014. Epub 
      2009 Jun 25.