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Suitability Of In-House Developed Pt-Pcr Fro The Detection And Serotyping Of Dengue Virus In Pakistan

By: Kashif Iqbal Sahibzada | Dr. Abu Saeed Hashmi.
Contributor(s): Dr. Aftab | Ms. Asma Waris.
Material type: materialTypeLabelBookPublisher: 2013Subject(s): Institute of Biochemistry & BiotechnologyDDC classification: 1551,T Dissertation note: Dengue Virus (DENV) belongs to the genus Flavivirus of family Flaviviridae having four serological different serotypes such as DENV1, DENV2, DENV3 and DENV4 (Bai et al., 2008) Being a Flaviviridae member, the dengue virus is transmitted to human by genus Aedes, mainly Aedes agypti. Over the years dengue fever has become a significant infectious disease in different parts of the world that leads and increases the growth of mosquitoes. It has become epidemic in more than 100 countries on the globe with more than 2.5 billion people at the risk of infection. Pakistan has witnessed some severe outbreaks of dengue viral infection which results to major morbidity and mortality since mid of 90s. There is a need to overcome this infectious and in many cases fatal disease. Imprecise fatality morbidity and statistics underrate the magnitude of dengue as a regional health problem. Medical and public health services have been incapable to diminish this infection since there is no current vaccine available to prevent infectious disease, no effective medical treatments that avert the development of severe symptoms and no sustainable control measures against the vector that guarantee protection of affected communities. Management of dengue patients and principally dengue hemorrhagic fever (DHF)/Dengue shock syndrome (DSS) cases are the alarming challenges now a day and in the upcoming episodes in this country. To deal with this challenge a sensitive and specific technique is required for its early diagnosis along with the knowledge of dengue serotype to increase the specificity of diagnosis and treatment. This study was designed to check the usefulness of nucleic acid based molecular determination of dengue virus along with nucleic acid sequencing/ analysis of different Dengue serotypes through phylogenetic studies. Total 50 Blood samples were collected from the dengue suspected patients in 2011 outbreak of dengue. Samples were analyzed by PCR based detection and were compared with IgG, IgM detections to check the usefulness of PCR based nucleic acid detection. In second phase of study nucleic acid sequencing was done The study has recommended PCR as a suitable and sensitive method for the rapid detection of dengue virus as it was found more sensitive than other utilized techniques including antibodies detection however it was not found useful to differentiate between primary and secondary infection for which a combination of IgG, IgM is more helpful choice. Nucleic acid analysis helped to define the common serotypes/genotypes of dengue virus circulating in Pakistan. In addition the present study has correlated our studied serotypes to other serotypes circulating in the globe which showed 98% homology with Srilankan strain and find out sequence similarities of our serotypes to the other serotypes distributed worldwide through phylogenetic analysis.
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Dengue Virus (DENV) belongs to the genus Flavivirus of family Flaviviridae having four serological different serotypes such as DENV1, DENV2, DENV3 and DENV4 (Bai et al., 2008) Being a Flaviviridae member, the dengue virus is transmitted to human by genus Aedes, mainly Aedes agypti. Over the years dengue fever has become a significant infectious disease in different parts of the world that leads and increases the growth of mosquitoes. It has become epidemic in more than 100 countries on the globe with more than 2.5 billion people at the risk of infection. Pakistan has witnessed some severe outbreaks of dengue viral infection which results to major morbidity and mortality since mid of 90s. There is a need to overcome this infectious and in many cases fatal disease. Imprecise fatality morbidity and statistics underrate the magnitude of dengue as a regional health problem. Medical and public health services have been incapable to diminish this infection since there is no current vaccine available to prevent infectious disease, no effective medical treatments that avert the development of severe symptoms and no sustainable control measures against the vector that guarantee protection of affected communities.
Management of dengue patients and principally dengue hemorrhagic fever (DHF)/Dengue shock syndrome (DSS) cases are the alarming challenges now a day and in the upcoming episodes in this country. To deal with this challenge a sensitive and specific technique is required for its early diagnosis along with the knowledge of dengue serotype to increase the specificity of diagnosis and treatment. This study was designed to check the usefulness of nucleic acid based molecular determination of dengue virus along with nucleic acid sequencing/ analysis of different Dengue serotypes through phylogenetic studies.
Total 50 Blood samples were collected from the dengue suspected patients in 2011 outbreak of dengue. Samples were analyzed by PCR based detection and were compared with IgG, IgM detections to check the usefulness of PCR based nucleic acid detection. In second phase of study nucleic acid sequencing was done
The study has recommended PCR as a suitable and sensitive method for the rapid detection of dengue virus as it was found more sensitive than other utilized techniques including antibodies detection however it was not found useful to differentiate between primary and secondary infection for which a combination of IgG, IgM is more helpful choice. Nucleic acid analysis helped to define the common serotypes/genotypes of dengue virus circulating in Pakistan. In addition the present study has correlated our studied serotypes to other serotypes circulating in the globe which showed 98% homology with Srilankan strain and find out sequence similarities of our serotypes to the other serotypes distributed worldwide through phylogenetic analysis.

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