Development of Molecular Diagnosis by PCR for the Detection of Infection and Gene Expression for Nipah Virus (NiV)
Autor: | Manal Amir, Farhina Pasha, Uzma Faridi, Aishah Alatawi |
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Rok vydání: | 2020 |
Předmět: |
Gene Expression Regulation
Viral biology medicine.diagnostic_test Nipah virus Nipah Virus Polymerase Chain Reaction Virology Virus law.invention Western blot law Gene expression biology.protein medicine Recombinant DNA Asian country Humans Pathology Molecular Antibody Primer (molecular biology) Agronomy and Crop Science |
Zdroj: | Pakistan Journal of Biological Sciences. 23:1086-1095 |
ISSN: | 1028-8880 |
Popis: | BACKGROUND AND OBJECTIVE The epidemiology of Nipah virus (NiV) was shortly seen in many Asian countries like Malaysia, Bangladesh and India most recently. Nipah virus also synonym as bat born virus is transmitted primarily by fruit bats. The 2 different strains transmitted are Hendra (highly pathogenic) and Cedar (non-pathogenic). The present study was attempt to develop recombinant protein based reagents for molecular diagnosis of Nipah. MATERIALS AND METHODS The different primer sets were developed using bioinformatics software DNASTAR. The E. coli cells were used for recombinant protein expression. RESULTS The NiV 'G' region primers were designed and amplified for 1 kb fragment and cloned. The NiV 'G' fragments were sub-cloned in pET-28(+) B and pGEX-5x-1. Recombinant protein thus obtained in soluble form in both the cases was essayed using western blot. The result showed the protein expression yield was more in pET-28(+) B with low stability and vice versa for pGEX-5x-1. CONCLUSION The antibodies raised from the protein can be used as diagnostic reagent for detection of NiV. Thus, a new diagnostic technique can be industrialized. |
Databáze: | OpenAIRE |
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