Hines RS, Wohlfarth GW, Moav R, Hulata G, 1974. Another challenge with regards to the vaccine is that no diagnostic tests are commercially available that can differentiate vaccinated versus naturally infected/exposed fish. CCB cells grown at 22°C were infected with wt CyHV-3. DNA was extracted from gill (panel A) and kidney (panel B) and analyzed by real-time TaqMan PCR for quantification of viral genome copies. Correction (i) is located in ORF51 and results in the deletion of an amino acid residue from the encoded protein. The membrane was prehybridized with prehybridization buffer (Roche Diagnostics, Indianapolis, IN) at 68°C, and then hybridized with the DIG-labeled DNA probe specific for ORF6 (Fig. 2005, Sano et al.
Specificity and sensitivity are the most important parameters to evaluate an assay. cDNA samples were further diluted 25 times in nuclease-free water prior to real-time quantitative PCR (RT-qPCR) analysis. Four CyHV-3 DNA sequences were analyzed using translating BLAST (http://www.ncbi.nlm.nih.gov/BLAST). This method involved synthesizing first-strand cDNA by reverse transcription by using a tagged, oligo(dT)-containing primer (thus contributing a 3′-PCR priming site) and a polymerase that adds a few nontemplated residues to the 3′ end of the cDNA. After three washes, samples were incubated at 25°C for 30 min with Alexa Fluor 568 goat anti-mouse immunoglobulin G (H+L) (GAM 568; 2 μg/ml; Molecular Probes) as the secondary conjugate. The length of the target fragments corresponds to the length in the sequence of CyHV-3 Japanese strain (accession no. Sensitivity of the nested PCR assay.
These results suggest that the virus carriers release CyHV-3 even in winter when viral activity is predicted to be at its lowest. Water temperature changes, which were measured by Uchii et al. Hence these proteins were called tegument proteins. However, these differences resulted in no alteration of the amino acid sequence (GenBank accession JQ815364), but only one amino acid alteration (GenBank accession EU349287). When the sequence alignment (BLAST) was performed, 99% nucleotide identity to the published CyHV-2 sequences (GenBank accession JQ815364, EU349287) was observed. RNA pellets were then collected by centrifugation, washed with 80% MBG EtOH, air dried and resuspended in 10 mM Tris-HCl pH 7.5. A few aspects of the interpretation were also strengthened by comparisons with homologous ORFs in the more distantly related IcHV-1 genome (7).
Jung ST, Miyazki T. Louis, USA), containing 10% (v/v) foetal calf serum (FCS) (Sigma-Aldrich), 2.5 mL penicillin streptomycin (PenStrep) 1250 units (U) (10000 U penicillin; 10 mg mL−1 streptomycin (Sigma-Aldrich)), 5 mL l-glutamine (200 mM) (Sigma-Aldrich) and 5 mL sodium pyruvate (100 mM) (Sigma-Aldrich). 2015; Nov 9; Online first. -+ Toxicological analysis of the water did not demonstrate any elevated heavy metals except for copper, measuring 0.02ppm (reference at 0.01ppm). (b) Water sampling locations at Ibanaiko. Three sets of blood samples from these six koi were collected to ensure the consistency of KHV genome detection as persistence of the genome over time is a characteristic of latency. An easier and faster extraction method than the use of the commercial kits for LAMP was examined.
determining the level, if any, of cross-protection that exposure to these viruses may confer on wild carp populations. 2011. 1. 2014). If specimens are frozen, they must remain frozen in transit and not be allowed to thaw out. The Bayesian maximum-likelihood tree was rooted by using bacteriophages T4 and RB69. Other fish species, like goldfish, and sturgeons, kept in the same ponds were not affected.
This tool should improve our knowledge regarding the present distribution and future diversification of this emerging virus. The presence of the KHV genome in IgM⁺ WBC was about 20-fold more abundant than in IgM-WBC. It can also enrich our understanding about the relationship between the CyHV-3 and its hosts by analysing their codon usage patterns. Sequence analyses of the enlarged 9/5, SphI-5 and TK gene regions identified both the Asian 1 (n = 11) and European 4 (n = 4) genotypes. During challenge experiments the peak of mortality caused by CyHV-3 was observed at days 8-12 p.i. S Corbeil, A Colling, LM Williams, FYK Wong, K Savin, S Warner, … Molecular Ecology Resources Link 山中裕樹, 源利文, 高原輝彦, 内井喜美子, 土居秀幸 (2016) 環境DNA分析の野外調査への展開.
It is closely related to CyHV-2 which attacks haematopoietic (blood cell producing) organs such as the liver, spleen, and kidneys of goldfish. Current diagnostics for detecting CyHV-3 are limited in sensitivity and are further complicated by latency. CyHV-3 ORF134 encodes an interleukin-10 (IL-10) homologue. L’utilisation de certaines versions du navigateur Internet Explorer 6 pose des problèmes d’affichage. Classification CyHV-3 is a member of the order Herpesvirales and newly designated family Alloherpesviridae (5,6) (Figure 2, panel A). Gli Alloherpesviridae sono una famiglia di virus facente parte dell’ordine Herpesvirales, nel gruppo I della Classificazione di Baltimore. There are no comments yet.
Cyprinid herpesvirus 3 (CyHV-3), previously designated carp interstitial nephritis and gill necrosis virus or koi herpesvirus, is the cause of a worldwide mortal disease of koi and carp. Copyright © 2014 Hui Zhang et al.