bAvian oncogenic viruses include Marek's disease virus (MDV), a highly contagious herpesvirus, as well as retroviruses such as avian leukosis virus (ALV) subgroups A to J and reticuloendotheliosis virus (REV). In this study, we examined the incidence of these viruses in suspected samples collected from poultry layer farms of South India, mainly in the Namakkal district of Tamil Nadu, a highly dense poultry-growing area in India. The histopathology-positive tissue sections were identified and further confirmed by immunohistochemistry using virus-specific antibodies. The viruses belonging to all 3 groups (MDV, ALV, and REV) were isolated in a cell culture system and confirmed by immunofluorescence using virus-specific antibodies. PCR appeared to be the method of choice for rapid and accurate diagnosis of these viruses. The multiplex PCR primers specific to MDV, ALV, REV, and chicken DNA were designed for rapid differential diagnosis. The specificity of the primers was checked by amplification of DNA from virus-infected cell culture in comparison with uninfected samples, and sensitivity was evaluated by calculating the minimum copy number at which amplification occurs in the cloned PCR products. The sequences of the amplicons were compared by BLAST analysis. PCR tests demonstrated the presence of single, dual, or triple viruses in some of the samples. Of 169 samples screened by multiplex PCR, 9 samples were positive for MDV, 17 samples were positive for ALV, 12 samples were positive for REV, and 17 samples were positive for both ALV and REV. Three samples were positive for all three viruses. ALV-positive samples were further subjected to subgroup-specific PCR, which gave positive results for subgroups B and D but not for subgroup J. Multiplex PCR appeared to be a useful technique for rapid differential diagnosis of avian oncogenic viruses and detection of multiple infections of avian oncogenic viruses under field conditions.
Methicillin-resistant Staphylococcus aureus (MRSA) is classified as hospital associated (HA), community associated (CA), livestock associated (LA) and is a global concern. Developing countries, like India, are densely populated country challenging for public hygiene practices. HA-MRSA is comfortably recorded in India, and CA-MRSA is also reported as increasing one. CA-MRSA is serious disease which affects the community as endemic. MRSA is one among major mastitis-causing organisms in India as LA-MRSA. There were reports for transmission of MRSA as community between milk handlers and cow in global perspective. In India reports of MRSA in short among milk handlers and also transmission between animal and human. Hence, proper monitoring of MRSA transmission in India should be elucidated in account among milk handlers and dairy cows to avoid emerging CA-MRSA as outbreak.
Summary. -Loop-mediated isothermal amplifi cation (LAMP) is known as a rapid and reliable alternative to conventional single-step or nested PCR for detection of genomic DNA of various pathogens in clinical samples. In this study, LAMP assay was developed for canine parvovirus (CPV) and compared with single-step and nested PCR assays. Out of 50 fecal samples from dogs clinically suspected for CPV infections, 19 were found positive by single-step PCR, 22 by nested PCR and 26 by LAMP. LAMP products were subjected to restriction analysis and sequencing to check their specifi city. LAMP assay turned out to be a rapid and fairly reproducible method, did not amplify other common canine pathogens and was more sensitive than nested PCR assay. Th erefore, it can be regarded as a highly reliable method for routine fi eld diagnosis of CPV infection.Keywords: canine parvovirus; nested polymerase chain reaction; loop-mediated isothermal amplifi cation; sensitivity; specifi city E-mail: drparthiban66@gmail.com; phone: +91-44-25369301. Abbreviations: CPV = canine parvovirus; LAMP = loop-mediated isothermal amplifi cation; LAT = latex agglutination test; MDCK = Madin Darby Canine Kidney; NS1, NS2 = non-structural protein 1 and 2; VP1, VP2 = viral protein 1 and 2
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