Dengue 1/2/3/4 Genotyping Detection Kit (Real-time RT-PCR)

Cat. No.: UP-ODEN-50T

[INTENDED USE]

Ultrassay Dengue genotyping detection kit is used for qualitative typing detection of denguevirus (DENV) nucleic acid in suspected patient’s serum sample to help diagnose patients with Dengue fever. The test results obtained by the kit are for clinical reference only. To make final diagnosis, comprehensive analysis of patient’s condition must be performed in conjunction with the patient’s clinical manifestations and other laboratory findings.

Dengue fever (DF), which is induced by denguevirus (DENV) infection, is one of the most epidemic arbovirus infectious diseases. Its transmission medium includes Aedes aegypti and Aedes albopictus. DF is mainly prevalent in tropical and subtropical areas [1]. DENV belongs to flavivirus under flaviviridae, and can be classified into 4 serotypes according to surface antigen. Clinical   manifestations of DENV infection mainly include headache, fever, weakness, enlargement of lymph node, leukopenia and etc., and bleeding, shock, hepatic injury or even death in severe cases [2]. In recent years, climate change, urbanization, quick development of tourism and other factors have provided more rapid and convenient conditions for transmitting and spreading of DF,  leading to constant expansion of epidemic area of DF [3]. Therefore, establishment of easy, specific and rapid etiological diagnosis method of DENV has important significance for clinical diagnosis of DF [4] .

[TEST PRINCIPLE]

This kit uses PCR combined with Taqman fluorescent probes. It contains specific primers and probes for fluorescence detection, which are designed to target the specific conserved regions of DENV I/II/III/IV. The  I/II/III/IV-specific probes are labeled with the fluorophore FAM/VIC (HEX)/ROX/CY5 at the 5′-end, and the quencher BHQ1 or BHQ2 at the 3’-end, respectively. The extracted RNA is reverse transcribed to cDNA. During PCR amplification, the specific primers and probes separately bind to their own target sequences. When the Taq DNA polymerase encounters the probes binding to their target sequences, it exhibits the exonuclease activity at the 5′-end to isolate the fluorophores (reporter dyes) from the quencher, thereby allowing the fluorescence-monitoring system to receive fluorescence signals, i.e., when a DNA strand is amplified, a fluorescent molecule will be formed to achieve complete  synchronization of PCR product formation and fluorescence signal accumulation, thereby qualitatively detecting DENV I/II/III/IV in a sample through 1 Reaction Mix.

[ACCEPTABLE SPECIMENS]

Fresh serum

[APPLICABLE EQUIPMENT]

Applicable to Ultrassay XP96/Mini 16 Real time qPCR system, ABI 7500 Real-Time PCR thermocycles, Bio-Rad CFX 96.