34 results about "Hantavirus" patented technology

The invention discloses a degenerate reverse transcription-polymerase chain reaction (RT-PCR) detection reagent and a kit for hantavirus group. The detection reagent comprises a pair of degenerate heterozygous oligonucleotide primers, and the sequences of the degenerate heterozygous oligonucleotide primers are respectively G1: GCAACAGCAACATGGTTTcartaytayac and G2: CTTCTTCATTCATATTTCCATGCarnccyttytc; the non-merger consensus sequence of the 5' ends in the primers plays a role in stabilizing the combination of a 3' merger core area and a template under the condition that the degeneracy of the primers is not increased, so that the specificity of the degenerate PCR reaction is improved, various viruses in hantavirus can be amplified and detected, the homologous unknown viruses of the extendedgenes can also be detected and the amplified target fragments can be sequenced by the detection reagent; and the kit are relatively high in sensitivity and good in hantavirus group specificity, can be used for detecting domestic popular Hantaan and Seoul viruses, and can also be used for detecting other oversea epidemic strains. The degenerate RT-PCR detection reagent and the kit for the hantavirus group are high in sensitivity, good in hantavirus specificity and universal.

The invention discloses a primer, kit and method for conducting genetic typing on the hantavirus by means of a PCR direct sequencing method. The primer comprises an upstream primer: ATTAGCCCWGTCATGAGTGT, and a downstream primer: CTTTGACTCYTTTGKYTCCA, wherein the Y is a C or a T, the W is an A or a T, and the K is a G or a T. The method comprises the steps of extracting a total RNA of a virus sample to be tested, conducting reverse transcription to obtain a cDNA, using the cDNA as a template, utilizing the primer for conducting PCR amplification, conducting sequencing on an amplified target fragment, constructing a phylogenetic tree by using corresponding fragments of known genotype representative strains as a reference on the basis of sequence information of the target fragment, and conducting the genetic typing on the virus sample to be tested according to the phylogenetic tree. When the primer is used, a specificity sequence of an S gene of the hantavirus can be obtained by only conducting one-time PCR amplification, and the genetic typing can be easily, conveniently and rapidly carried out on the hantavirus by utilizing the specificity sequence.