AffiPCR® TBEV, B.burgdorferi, A.phagocytophilum, E.chaffeensis / E.muris Real Time PCR

Target Disease Type: Dangerous Infections

Specific Application: Tick-Borne Encephalitis

Storage and Shipping : 4 weeks

TBEV, B.burgdorferi, A.phagocytophilum, E.chaffeensis / E.muris RT-PCR (CE) V59-100FRT DataSheet

INTENDED USE

TBEV, B.burgdorferi sl, A.phagocytophilum, E.chaffeensis / E.muris Real-TM PCR kit is an in vitro nucleic acid amplification test for qualitative detection of RNA of tick-borne encephalitis virus (TBEV), Borrelia burgdorferi sl (Ixodes tick-borne borreliosis (ITB) pathogen), Ehrlichia chaffeensis andEhrlichia muris (human monocytic ehrlichiosis (HME) pathogens) and DNA of Anaplasma phagocytophilum (human granulocytic anaplasmosis (HGA) pathogen) in biological materials (ticks, blood, cerebrospinal fluid, and autopsy material) by using real-time hybridization-fluorescence detection.

PRINCIPLE OF ASSAY

TBEV, B.burgdorferi sl, A.phagocytophilum, E.chaffeensis / E.muris nucleic acid detection by the polymerase chain reaction (PCR) is based on the amplification of pathogen genome specific region using specific primers. In real-time PCR, the amplified product is detected using fluorescent dyes. These dyes are linked to oligonucleotide probes which bind specifically to the amplified product during thermocycling. The real-time monitoring of fluorescence intensities during the real-time PCR allows the detection of accumulating product without re-opening the reaction tubes after the PCR run. TBEV, B.burgdorferi sl, A.phagocytophilum, E.chaffeensis / E.muris Real-TM PCR kit is a qualitative test that contains the Internal Control (IC) . It must be used in the extraction procedure in order to control the extraction process of each individual sample and to identify possible reaction inhibition. TBEV, B.burgdorferi sl, A.phagocytophilum, E.chaffeensis / E.muris Real-TM PCR kit uses “hot-start”, which greatly reduces the frequency of nonspecifically primed reactions. “Hot-start” is guaranteed by separation of nucleotides and Taq-polymerase by using a chemically modified polymerase (TaqF) . Chemically modified polymerase (TaqF) is activated by heating at 95 °C for 15 min.

TBEV, B.burgdorferi sl, A.phagocytophilum, E.chaffeensis / E.muris nucleic acid detection includes:

1. RNA/DNA extraction from biological material simultaneously with the Internal Control;
2. reverse transcription of cDNA on RNA template;
3. PCR with real-time detection of cDNA/DNA amplification products.

STABILITY

TBEV, B.burgdorferi sl, A.phagocytophilum, E.chaffeensis / E.muris Real-TM Test is stable up to the expiration date indicated on the kit label. The product will maintain performance through the control date printed on the label. The shelf life of reagents before and after the first use is the same, unless otherwise stated. Exposure to light, heat or humidity may affect the shelf life of some of the kit components and should be avoided. Repeated thawing and freezing of these reagents should be avoided, as this may reduce the sensitivity. Components stored under conditions other than those stated on the labels may not perform properly and may adversely affect the assay results.