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Molecular Identification Of Soil Borne Bacillus Anthracis From Districts Lahore And Sheikhupura

By: Tariq Jamil | Prof. Dr. Masood Rabbani.
Contributor(s): Dr. Muhammad Zubair Shabbir | Prof. Dr.
Material type: materialTypeLabelBookPublisher: 2013Subject(s): Department of MicrobiologyDDC classification: 1817,T Dissertation note: Background: Anthrax is a bacterial zoonotic disease caused by Bacillus anthracis. Accurate assays for etiological identification are necessary to ensure proper veterinary and medical health facilities against such diseases. Real-time PCR is a powerful technique to identify this organism based on the presence of two unique plasmids (pXO1 and pXO2) and is highly preferable technique over conventional detection assays in clinical and environmental samples both. Methodology: Real Time-PCR technique was used to identify Bacillus anthracis bacteria in the soils of districts Lahore and Sheikhupura. Soil samples were collected from each village of both districts and processed for genome extraction using commercial soil DNA extraction kit. Following genome extraction, the samples were run further for real-time PCR analysis. Positive controls, primers and probes were provided by the Penn state University. SPSS software and pearson's chi square distribution test were used for statistical analysis. Findings and Suggestions: Real-time PCR was found as a powerful tool to detect Bacillus anthracis in environmental samples. The bacterium detected was of non-virulent type and showed associations with soil humidity and land use. Further studies may include study of the bacterium with respect to soil-chemistry and sero-prevalence among positive areas of the two districts. Strain characterization is also recommended. The present results may also help in ecological niche modelling by using spatial mapping techniques. Such studies will help in a better understanding of soil as a reservoir for zoonotic organisms and surveillance of the diseases.
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Background:
Anthrax is a bacterial zoonotic disease caused by Bacillus anthracis. Accurate assays for etiological identification are necessary to ensure proper veterinary and medical health facilities against such diseases. Real-time PCR is a powerful technique to identify this organism based on the presence of two unique plasmids (pXO1 and pXO2) and is highly preferable technique over conventional detection assays in clinical and environmental samples both.
Methodology:
Real Time-PCR technique was used to identify Bacillus anthracis bacteria in the soils of districts Lahore and Sheikhupura. Soil samples were collected from each village of both districts and processed for genome extraction using commercial soil DNA extraction kit. Following genome extraction, the samples were run further for real-time PCR analysis. Positive controls, primers and probes were provided by the Penn state University. SPSS software and pearson's chi square distribution test were used for statistical analysis.
Findings and Suggestions:
Real-time PCR was found as a powerful tool to detect Bacillus anthracis in environmental samples. The bacterium detected was of non-virulent type and showed associations with soil humidity and land use. Further studies may include study of the bacterium with respect to soil-chemistry and sero-prevalence among positive areas of the two districts. Strain characterization is also recommended. The present results may also help in ecological niche modelling by using spatial mapping techniques. Such studies will help in a better understanding of soil as a reservoir for zoonotic organisms and surveillance of the diseases.

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