Rapid and sensitive detection of Feline immunodeficiency virus using an insulated isothermal PCR-based assay with a point-of-need PCR detection platform.

摘要: Feline immunodeficiency virus (FIV) is an important infectious agent of cats. Clinical syndromes resulting from FIV infection include immunodeficiency, opportunistic infections, and neoplasia. In our study, a 5' long terminal repeat/gag region-based reverse transcription insulated isothermal polymerase chain reaction (RT-iiPCR) was developed to amplify all known strains facilitate point-of-need diagnosis. The RT-iiPCR method applied in PCR detection platform--a field-deployable device capable generating automatically interpreted results nucleic acids within 1 hr. Limit 95% calculated be 95 copies standard vitro RNA per reaction. Endpoint dilution studies with serial dilutions ATCC type strain showed that the sensitivity lyophilized reagent comparable reference nested PCR. established did not any nontargeted feline pathogens, including Felid herpesvirus 1, coronavirus, calicivirus, leukemia virus, Mycoplasma haemofelis, Chlamydophila felis. Based on analysis 76 clinical samples (including blood bone marrow) RT-iiPCR, test 97.78% (44/45), specificity 100.00% (31/31), agreement 98.65% (75/76), determined against nested-PCR assay. A kappa value 0.97 indicated excellent correlation between these 2 methods. reagent, deployed user-friendly portable device, has potential utility for rapid easy