One-step RT-PCR for the Detection of Infectious Bursal Disease Virus in Clinical Samples     

Kataria RS  Tiwari AK  Nanthakumar T  Goswami PP 《Veterinary research communications》2001,25(5):429-436

A single-tube, non-interrupted, one-step RT-PCR has been standardized to amplify the hypervariable region of the VP2 gene sequence of infectious bursal disease virus (IBDV). The technique standardized on purified viral RNA was successfully applied to the detection of the virus directly in clinical samples. The amplified products were confirmed to be IBDV specific by their size in ethidium bromide-stained agarose gel, nested PCR and restriction enzyme digestion. Digestion of the amplicons with StyI restriction enzyme also differentiated classical virus from six very virulent field isolates. The sensitivity of the one-step RT-PCR was found to be 0.2 pg of viral RNA.  相似文献   

Differential detection of Newcastle disease virus strains by degenerate primers based RT-PCR   总被引：2，自引：0，他引：2  

Tiwari AK  Kataria RS  Nanthakumar T  Dash BB  Desai G 《Comparative immunology, microbiology and infectious diseases》2004,27(3):163-169

Degenerate primers based RT-PCR (previously described by [Avian Dis 26 (1997) 837]) has been used for the detection and differentiation of Newcastle disease (ND) viruses. Two sets of primers (A+B and A+C), with common forward primer and distinct reverse degenerate primers, designed from fusion protein gene encoding for cleavage site, could differentiate virulent and avirulent Newcastle disease viruses (NDV). Both sets of primers amplified "F" gene sequence of virulent (velogenic and mesogenic) viruses, whereas in avirulent strains, amplification was only with primer set A+C. Total 10 NDV isolates and two clinical samples including both known and unknown pathotypes, were checked. Based on amplification results 5 viruses were found to be virulent type and 6 as avirulent with one of the two clinical samples, earlier positive by RT-PCR using non-degenerate "F" gene specific primers was found negative in this study. The technique has been found to be a simple and quick for the detection and differentiation of virulent and avirulent NDV, which is important for control of the disease in the events of the outbreaks.  相似文献   

Cloning and expression of a cocaine-sensitive dopamine transporter complementary DNA   总被引：41，自引：0，他引：41  

S Shimada  S Kitayama  C L Lin  A Patel  E Nanthakumar  P Gregor  M Kuhar  G Uhl 《Science (New York, N.Y.)》1991,254(5031):576-578

A rat dopamine (DA) transporter complementary DNA has been isolated with combined complementary DNA homology and expression approaches. The DA transporter is a 619-amino acid protein with 12 hydrophobic putative membrane-spanning domains and homology to the norepinephrine and gamma-aminobutyric acid transporters. The expressed complementary DNA confers transport of [3H]DA in Xenopus oocytes and in COS cells. Binding of the cocaine analog [3H]CFT ([3H]2 beta-carbomethoxy-3 beta-(4-fluorophenyl)tropane) to transfected COS cell membranes yields a pharmacological profile similar to that in striatal membranes.  相似文献   

Characterization of fowl adenovirus serotype-4 associated with hydropericardium syndrome in chicken     

Balamurugan V  Kataria JM  Kataria RS  Verma KC  Nanthakumar T 《Comparative immunology, microbiology and infectious diseases》2002,25(3):139-147

The polypeptides of three fowl adenovirus-4 (FAV-4) field isolates of hydropericardium syndrome from various geographical areas of the country and the standard FAV-1 (CELO virus) were subjected to sodium dodecyl sulfate polyacrylamide gel electrophoresis and analysed by protein immunoblotting with polyclonal antibodies to FAV-4 and FAV-1. Protein profile analysis of FAV-4 isolates revealed similarity of all the eight polypeptides with molecular weight ranging from 20 to 107 kDa but differed from CELO, particularly in their 24.2 kDa protein. Subsequent immunoblotting showed relatedness of at least five protein fractions of FAV-4 to CELO virus.  相似文献   

Pathotyping of Newcastle Disease Viruses by RT-PCR and Restriction Enzyme Analysis   总被引：3，自引：0，他引：3  

Nanthakumar T  Kataria RS  Tiwari AK  Butchaiah G  Kataria JM 《Veterinary research communications》2000,24(4):275-286

The technique of RT-PCR and restriction enzyme analysis was standardized to detect and differentiate Newcastle disease viruses. Digestion of RT-PCR-amplified, F gene sequences encoding for the cleavage activation sites of fusion protein with restriction enzymes AluI, BglI, HaeIII, HinfI, HhaI, RsaI, StyI and TaqI was carried out in order to characterize Newcastle disease viruses of varying pathogenicity. Restriction enzyme digestion of the amplicons by BglI and HhaI could group eight viruses, both field isolates and known vaccine strains, into lentogenic, mesogenic and velogenic pathotypes. By employing this technique directly on a clinical sample, Newcastle disease virus of the lentogenic pathotype could be detected.  相似文献