Abstract :
Background: Dengue is the most important viral mutagenic disease in public health, caused by one of the four
serotypes (DEN-1, 2, 3 and 4) of the dengue virus, a positive chain RNA virus of the Flaviviridae family, which
produces a spectrum of illness ranging from dengue fever to dengue hemorrhagic fever / shock syndrome (shock)
due to dengue, the latter a serious infection with vascular and haemostatic abnormality that can lead to death.
Objectives: To identify by reverse transcription-polymerase chain reaction (RT-PCR) and specific restriction sites -
polymerase chain reaction (RSS-PCR) to the causative agent of the epidemic outbreak presented in the district of
Faisalabad in April 2017.
Materials and methods: twenty serum samples collected during the dengue outbreak were processed by RT-PCR to
determine the serotype; this technique was performed in one step. The RSS-PCR technique was then applied to
identify the circulating genotype and the results were subsequently corroborated with viral isolation and
sequencing.
Results: The analysis of the RTPCR of the RNA extracted from the samples presented an amplified product of 290pb
corresponding to the dengue serotype. The analysis of the RSS-PCR products of RNA extracted from dengue isolates
corresponded to pattern C, included in genotype III. The isolations of the dengue virus in C6 / 36 cell lines, typed by
IFI and the genetic sequencing confirmed the results obtained by the tests previously described. Conclusion: During
the classic dengue epidemic outbreak in Lima, genotype III of the dengue virus circulated.
Keywords: Classic dengue outbreak; dengue virus; molecular typing; RT-PCR; RSS-PCR.
Keyword :
Classic dengue outbreak; dengue virus; molecular typing; RT-PCR; RSS-PCR.