Loop-mediated isothermal amplification for rapid detection of Chinese sacbrood virus

Ma, Mai‐Juan; Ma, Chao; Li, Mng; Wang, Shuquan; Yang, Song; Wang, Shude

doi:10.1016/j.jviromet.2011.05.028

Cited by 24 publications

(9 citation statements)

References 21 publications

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“…Taking together phylogenetic data for SBV worldwide showed high diversity split in different clades from the Pacific region (Australia, China, South Korea, India, Vietnam, etc.) ( Roberts, Anderson & Durr, 2017 ; Ma et al, 2011 ; Xia, Zhou & Wei, 2015 ; Reddy et al, 2016 ; Nguyen & Le, 2013 ).…”

Section: Discussionmentioning

confidence: 99%

Molecular detection and phylogenetic assessment of six honeybee viruses in Apis mellifera L. colonies in Bulgaria

Shumkova

Neov

Sirakova

et al. 2018

PeerJ

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Honey bee colonies suffer from various pathogens, including honey bee viruses. About 24 viruses have been reported so far. However, six of them are considered to cause severe infection which inflicts heavy losses on beekeeping. The aim of this study was to investigate incidence of six honey bee viruses: deformed wing virus (DWV), acute bee paralysis virus (ABPV), chronic bee paralysis virus (CBPV), sacbrood virus (SBV), kashmir bee virus (KBV), and black queen cell virus (BQCV) by a reverse transcription polymerase chain reaction (RT-PCR). A total of 250 adult honey bee samples were obtained from 50 colonies from eight apiaries situated in three different parts of the country (South, North and West Bulgaria). The results showed the highest prevalence of DWV followed by SBV and ABPV, and one case of BQCV. A comparison with homology sequences available in GenBank was performed by phylogenetic analysis, and phylogenetic relationships were discussed in the context of newly described genotypes in the uninvestigated South Eastern region of Europe. In conclusion, the present study has been the first to provide sequencing data and phylogenetics analyses of some honey bee viruses in Bulgaria.

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Section: Discussionmentioning

confidence: 99%

Molecular detection and phylogenetic assessment of six honeybee viruses in Apis mellifera L. colonies in Bulgaria

Shumkova

Neov

Sirakova

et al. 2018

PeerJ

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“…A total of 50 infected A. mellifera larvae displaying upward warping of the head and body surface changes, and white or sick and dead larvae in the honeycomb with the capped brood abnormally sealed, were collected, weighed, and completely homogenized in sterile water (1.5× by weight) using a mortar and pestle. AmCSBV-SDLY-2016 purification was performed by cesium chloride gradient centrifugation, according to the method reported by Ma et al (2011b, 2011c). The supernatant was extracted with an equal volume of 1,1,2-trichlorotrifluoroethane before the aqueous phase was layered over a discontinuous CsCl gradient (1.5 and 1.2 g/cm 3 ) and centrifuged at 270,000 ×g for 1 h. The material at the CsCl interface was harvested and adjusted to a volume of five mL (final density 1.38 g/cm 3 ) with CsCl solution and centrifuged at 270,000 ×g overnight.…”

Section: Methodsmentioning

confidence: 99%

Genetic and phylogenetic analysis of Chinese sacbrood virus isolates fromApis mellifera

Li¹,

Fei²,

Sun³

et al. 2019

PeerJ

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BackgroundSacbrood virus (SBV) is one of the most pathogenic honeybee viruses that exhibits host specificity and regional variations. The SBV strains that infect the Chinese honeybee Apis cerana are called Chinese SBVs (CSBVs).MethodsIn this study, a CSBV strain named AmCSBV-SDLY-2016 (GenBank accession No. ) infecting A. mellifera was identified by electron microscopy, its protein composition was analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis and agar gel immunodiffusion assay, and its nucleotide sequence was identified using a series of reverse-transcription polymerase chain reaction fragments of AmCSBV-SDLY-2016 generated using SBV/CSBV-specific primers. To investigate phylogenetic relationships of the CSBV isolates, a phylogenetic tree of the complete open reading frames (ORF) of the CSBV sequences was constructed using MEGA 6.0; then, the similarity and recombination events among the isolated CSBV strains were analyzed using SimPlot and RDP4 software, respectively.ResultsSequencing results revealed the complete 8,794-nucleotide long complete genomic RNA of the strain, with a single large ORF (189–8,717) encoding 2,843 amino acids. Comparison of the deduced amino acid sequence with the SBV/CSBV reference sequences deposited in the GenBank database identified helicase, protease, and RNA-dependent RNA polymerase domains; the structural genes were located at the 5′ end, whereas the non-structural genes were found at the 3′ end. Multiple sequence alignment showed that AmCSBV-SDLY-2016 had a 17-amino acid (aa) and a single aa deletion at positions 711–729 and 2,128, respectively, as compared with CSBV-GD-2002, and a 16-aa deletion (positions 711–713 and 715–728) as compared with AmSBV-UK-2000. However, AmCSBV-SDLY-2016 was similar to the CSBV-JLCBS-2014 strain, which infects A. cerana. AmCSBV-SDLY-2016 ORF shared 92.4–97.1% identity with the genomes of other CSBV strains (94.5–97.7% identity for deduced amino acids). AmCSBV-SDLY-2016 was least similar (89.5–90.4% identity) to other SBVs but showed maximum similarity with the previously reported CSBV-FZ-2014 strain. The phylogenetic tree constructed from AmCSBV-SDLY-2016 and 43 previously reported SBV/CSBV sequences indicated that SBV/CSBV strains clustered according to the host species and country of origin; AmCSBV-SDLY-2016 clustered with other previously reported Chinese and Asian strains (AC genotype SBV, as these strains originated from A. cerana) but was separate from the SBV genomes originating from Europe (AM genotype SBV, originating from A. mellifera). A SimPlot graph of SBV genomes confirmed the high variability, especially between the AC genotype SBV and AM genotype SBV. This genomic diversity may reflect the adaptation of SBV to specific hosts, ability of CSBV to cross the species barrier, and the spatial distances that separate CSBVs from other SBVs.

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“…After weighing, larvae were completely homogenised in sterile water (1.5-fold amount, by weight) using a pestle and mortar. CSBV purification was performed by cesium chloride gradient centrifugation, according to Ma’s method1232. The supernatant was then passed through a 0.45-μm cell filter first and then through a 0.22-μm cell filter.…”

Section: Methodsmentioning

confidence: 99%

A comparison of biological characteristics of three strains of Chinese sacbrood virus in Apis cerana

Fei

Jiang

et al. 2016

Sci Rep

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We selected and sequenced the entire genomes of three strains of Chinese sacbrood virus (CSBV): LNQY-2008 (isolated in Qingyuan, Liaoning Province), SXYL-2015 (isolated in Yulin, Shanxi Province), and JLCBS-2014 (isolated in Changbaishan, Jilin Province), by VP1 amino acid (aa) analysis. These strains are endemic in China and infect Apis cerana. Nucleotide sequences, deduced amino acid sequences, genetic backgrounds, and other molecular biological characteristics were analysed. We also examined sensitivity of these virus strains to temperature, pH, and organic solvents, as well as to other physicochemical properties. On the basis of these observations, we compared pathogenicity and tested cross-immunogenicity and protective immunity, using antisera raised against each of the three strains. Our results showed that compared with SXYL-2015, LNQY-2008 has a 10-aa deletion and 3-aa deletion (positions 282–291 and 299–301, respectively), whereas JLCBS-2014 has a 17-aa deletion (positions 284–300). However, the three strains showed no obvious differences in physicochemical properties or pathogenicity. Moreover, there was immune cross-reactivity among the antisera raised against the different strains, implying good protective effects of such antisera. The present study should significantly advance the understanding of the pathogenesis of Chinese sacbrood disease, and offers insights into comprehensive prevention and treatment of, as well as possible protection from, the disease by means of an antiserum.

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Loop-mediated isothermal amplification for rapid detection of Chinese sacbrood virus

Cited by 24 publications

References 21 publications

Molecular detection and phylogenetic assessment of six honeybee viruses in Apis mellifera L. colonies in Bulgaria

Molecular detection and phylogenetic assessment of six honeybee viruses in Apis mellifera L. colonies in Bulgaria

Genetic and phylogenetic analysis of Chinese sacbrood virus isolates fromApis mellifera

A comparison of biological characteristics of three strains of Chinese sacbrood virus in Apis cerana

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