Development And Optimization Of Multiplex Pcr For The Identification Of A, O And Asia 1 Strains Of FMDV In Pakistan
By: Muhammad Ikram
| Dr. Atif Hanif.
Contributor(s): Dr. Imran Najeeb | Prof. Dr.
Material type: 
BookPublisher: 2010Subject(s): Department of MicrobiologyDDC classification: 1189,T Dissertation note: Foot and mouth disease (FMD) is highly infectious disease of cattle, buffalo, sheep and goats. It is caused by genus Aphthovirus of Picomaviradae family. FMDV is RNA virus having seven serotypes A, 0, C, Asia I, SAT1, SAT2 and SAT3.
Foot and mouth disease is endemic in Pakistan and causes high economic losses to livestock industry. So priority is to develop quick and efficient methods for detection of FMDV and to limit the spread of disease outbreak. Although CFT, VNT and ELISA are already being used for the diagnosis of FMDV in Pakistan but these diagnostic techniques are time consuming and their specificity and sensitivity is low. Multiplex PCR for the identification of FMDV is very much sensitive and specific, can be done with in three hours after the receipt of samples.
Present study has been designed to optimize multiplex RT-PCR for rapid detection of FMD virus. RNA was extracted from virus stock obtained from QOL, UVAS Lahore and from field samples. After RNA extraction the samples were subjected to synthesize cDNA by the use of Reverse Transcriptase enzyme. After cDNA synthesis PCR reaction was carried out. The amplified products were resolved on 1.5% Agarose Gel. A multiplex RT-PCR strategy was optimized and developed for the detection of virus serotypes A, 0 and Asia l.
Restulst of this study helped to develop an efficient and economical method for rapid detection of FMD virus and also helpful in differential diagnosis from other vesicular diseases.
| Item type | Current location | Collection | Call number | Status | Date due | Barcode | Item holds |
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Thesis
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UVAS Library Thesis Section | Veterinary Science | 1189,T (Browse shelf) | Available | 1189,T |
Foot and mouth disease (FMD) is highly infectious disease of cattle, buffalo, sheep and goats. It is caused by genus Aphthovirus of Picomaviradae family. FMDV is RNA virus having seven serotypes A, 0, C, Asia I, SAT1, SAT2 and SAT3.
Foot and mouth disease is endemic in Pakistan and causes high economic losses to livestock industry. So priority is to develop quick and efficient methods for detection of FMDV and to limit the spread of disease outbreak. Although CFT, VNT and ELISA are already being used for the diagnosis of FMDV in Pakistan but these diagnostic techniques are time consuming and their specificity and sensitivity is low. Multiplex PCR for the identification of FMDV is very much sensitive and specific, can be done with in three hours after the receipt of samples.
Present study has been designed to optimize multiplex RT-PCR for rapid detection of FMD virus. RNA was extracted from virus stock obtained from QOL, UVAS Lahore and from field samples. After RNA extraction the samples were subjected to synthesize cDNA by the use of Reverse Transcriptase enzyme. After cDNA synthesis PCR reaction was carried out. The amplified products were resolved on 1.5% Agarose Gel. A multiplex RT-PCR strategy was optimized and developed for the detection of virus serotypes A, 0 and Asia l.
Restulst of this study helped to develop an efficient and economical method for rapid detection of FMD virus and also helpful in differential diagnosis from other vesicular diseases.
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