Development of non radioactive DNA probe for diagnosis of bovine herpes virus-1
Abstract
Bovine herpesvirus-l (BHV-1), a member of the family Herpesviridae and subfamily Alphaherpesvirinae is an important cause of respiratory and genital diseases in cattle. Infection with BHV-1 occurs worldwide and causes serious economic losses due to mortality, abortions, decreased milk production and loss of weight. The current techniques for detection of BHV- 1 in the diagnostic laboratories include virus isolation, and antigen detection by ELISA. These techniques lack sensitivity and are both time consuming and expensive. Keeping this point in view the present study was undertaken to develop and evaluate the non radioactive probe for detection of BHV-1. The non radioactive DNA probe was successfully evaluated for detection of BHV-1 in biological field samples. By using DNA probe total a 4 Nasal swabs, 8 vaginal swabs, lung, liver and cotyledons of aborted material were detected positive. Efficiency of two different primer pairs belonging to gI and gB glycoproteins, for detection of BHV-1 was compared. The gI specific PCR detected upto 1fg of viral DNA from purified BHV-1. Whereas the gB gene specific PCR detected upto 10fg of nucleic acid
The gI gene specific PCR was used to confirm the results of probe hybridization assay. Probe positive all the samples also detected positive by PCR. None of the probe
negative samples was found positive by PCR. The techniques could be useful for screening of large number of biological samples. The chelex method of DNA isolation from clinical samples included a fast, safe and convenient extraction procedure. The study clearly indicated that, the novel extraction method with non radioactive DNA probe hybridization assay provide simple, rapid and sensitive diagnostic tool for detection of BHV-1 infection in biological samples especially when large number have to be processed.