ABSTRACT
Bluetongue virus (BTV) infects all ruminants, including cattle, goats and camelids, causing bluetongue disease (BT) that is often severe in naïve deer and sheep. Reverse-transcription-loop-mediated-isothermal-amplification (RT-LAMP) assays were developed to detect eastern or western topotype of BTV strains circulating in India. Each assay uses four primers recognizing six distinct sequences of BTV genome-segment 1 (Seg-1). The eastern (e)RT-LAMP and western (w)RT-LAMP assay detected BTV RNA in all positive isolates that were tested (n=52, including Indian BTV-1, -2, -3, -5, -9, -10, -16, -21 -23, and -24 strains) with high specificity and efficiency. The analytical sensitivity of the RT-LAMP assays is comparable to real-time RT-PCR, but higher than conventional RT-PCR. The accelerated eRT-LAMP and wRT-LAMP assays generated detectable levels of amplified DNA, down to 0.216 fg of BTV RNA template or 108 fg of BTV RNA template within 60-90min respectively. The assays gave negative results with RNA from foot-and-mouth-disease virus (FMDV), peste des petits ruminants virus (PPRV), or DNA from Capripox viruses and Orf virus (n=10), all of which can cause clinical signs similar to BT. Both RT-LAMP assays did not show any cross-reaction among themselves. The assays are rapid, easy to perform, could be adapted as a 'penside' test making them suitable for 'front-line' diagnosis, helping to identify and contain field outbreaks of BTV.
Subject(s)
Bluetongue virus/isolation & purification , Bluetongue/diagnosis , Goat Diseases/diagnosis , Nucleic Acid Amplification Techniques/methods , Real-Time Polymerase Chain Reaction/methods , Animals , Bluetongue/virology , Bluetongue virus/classification , Bluetongue virus/genetics , Cattle , DNA Primers , Genome, Viral , Goat Diseases/virology , Goats/virology , India , Nucleic Acid Amplification Techniques/standards , Peste-des-Petits-Ruminants/diagnosis , Peste-des-Petits-Ruminants/virology , Peste-des-petits-ruminants virus/genetics , Peste-des-petits-ruminants virus/isolation & purification , Real-Time Polymerase Chain Reaction/standards , Sensitivity and Specificity , Sheep/virology , TemperatureABSTRACT
Objective To study the effect of homeobox genes CDX1 and CDX2 on the phenotype of esophageal adenocarcinoma cells.Methods Esophageal adenocarcinoma cell line SEG-1 was transfected with CDX1 or CDX2 cDNA.The morphology,growth rate,division index and tumorigenicity were analyzed.Results The expression of CDX1 or CDX2 leaded to occurring adeniform-like manifestation.The growth rate,division index and tumorigenicity were reduced,especially by CDX2.Conclusion CDX1 and CDX2 all could increase differentiation of esophageal adenocarcinoma cells and reduction of its malignancy.
