Dengue-CHIK Duplex RT-PCR
Dengue (DEN) and Chikungunya (CHIK) have emerged as two most important arboviral infections of global significance. The similarities in clinical presentations, their circulation in the same geographical area and transmission through same vector necessitate an urgent need for the differential diagnosis of these two infections. The incidence of dual infection in the same individual further complicates the task of the clinicians to arrive at a definitive diagnosis. In the absence of any specific therapy or vaccine, early diagnosis is crucial for patient management.
This assay is based on the principle of duplex RT-PCR, by targeting genomic regions of both dengue and chikungunya virus. This assay is developed as a one step method by combining both RT and PCR steps. Further both primers are included in the single tube, making it a one-step, single tube protocol.
The presence of 511 bp indicates dengue complex and of 205 bp indicates Chikungunya infection. Presence of both amplicons suggests the dual infection in the same patient.
The evaluation was carried out with more than 400 clinical samples from recent chikungunya and dengue outbreaks in India. This assay could also able to detect dual infection of CHIK and DEN in patients. This kit has potential clinical and epidemiological application for rapid, sensitive detection, differentiation and genotyping of DEN and CHIK viruses in clinical samples.
The innovation is a testing kit for detection of Dengue (DEN) and chikungunya (CHIK) infection with high degree of sensitivity and specificity in a single clinical sample. Dengue and CHIKV is an insect-borne virus, of the genus, Alphavirus, that is transmitted to humans by virus-carrying mosquitoes in a human subject. The present innovation is based on the principle of duplex RT-PCR (Reverse Transcriptase Polymerase Chain Reaction) assay. The assay includes one-step method of combining both RT and PCR steps and targeting genomic strands of both Dengue and chikungunya virus. The presence of 511 bp indicates dengue complex and of 205 bp indicates Chikungunya infection. Presence of both amplicons suggests the dual infection in the same patient. There are various antiviral agents used against Chikungunya and Dengue infections but these antiviral agents are not effective. Further, till now there is no effective prophylaxis treatment available for Chikungunya and Dengue. The present innovation is better than the existing solution as it can detect the presence of Dengue and CHIKV virus in a single sample i.e. dual infection in the same patient. This assay could also able to detect dual infection of CHIK and DEN in patients. This kit has potential clinical and epidemiological application for rapid, sensitive detection, differentiation and genotyping of DEN and CHIK viruses in clinical samples. The kit will be useful in public health sector for early clinical diagnosis and surveillance of Dengue and Chikungunya infection.