I-Clostridium difficile toxin A/B gene (C.diff)
Igama lomkhiqizo
Ikhithi Yokuthola I-HWTS-OT031A Nucleic Acid ye-Clostridium difficile toxin A/B gene (C.diff) (Fluorescence PCR)
Isitifiketi
CE
I-Epidemiology
I-Clostridium difficile (CD), i-gram-positive anaerobic sporogenic Clostridium difficile, ingenye yezifo ezibangela ukutheleleka kwamathumbu e-nosocomial. Ngokwezokwelapha, cishe u-15% ~ 25% wesifo sohudo esihlobene namagciwane, u-50% ~ 75% we-antimicrobial-related colitis kanye no-95% ~ 100% we-pseudomembranous enteritis kubangelwa ukutheleleka kwe-Clostridium difficile (CDI). I-Clostridium difficile iyi-pathogen enemibandela, kufaka phakathi izinhlobo ze-toxigenic kanye nezinhlobo ezingezona ze-toxigenic.
Isiteshi
| I-FAM | i-tcdAi-gene |
| I-ROX | i-tcdBi-gene |
| I-VIC/HEX | Ukulawula Kwangaphakathi |
Amapharamitha Obuchwepheshe
| Isitoreji | ≤-18℃ |
| Isikhathi sokuphelelwa yisikhathi | Izinyanga ezingu-12 |
| Uhlobo Lwesifanekiso | isihlalo |
| Tt | ≤38 |
| CV | ≤5.0% |
| I-LoD | 200CFU/mL |
| Ukucaciswa | Sebenzisa le khithi ukuthola ezinye izifo zamathumbu ezifana ne-Escherichia coli, i-Staphylococcus aureus, i-Shigella, i-Salmonella, i-Vibrio parahaemolyticus, i-Group B Streptococcus, izinhlobo ze-Clostridium difficile ezingezona ezibangela izifo, i-Adenovirus, i-rotavirus, i-norovirus, i-influenza A virus, i-influenza B virus kanye ne-human genomic DNA, imiphumela yonke ayimibi. |
| Izinsimbi Ezisebenzayo | Izinhlelo ze-PCR ze-Applied Biosystems 7500 Real-Time Izinhlelo ze-PCR ze-Applied Biosystems 7500 Fast Real-Time I-QuantStudio®Izinhlelo ze-PCR zesikhathi sangempela ezi-5 Izinhlelo ze-PCR ze-SLAN-96P Zesikhathi Sangempela (i-Hongshi Medical Technology Co., Ltd.) I-LightCycler®Uhlelo lwe-PCR lwesikhathi sangempela lwe-480 Uhlelo Lokuthola i-LineGene 9600 Plus Real-Time PCR (I-FQD-96A，I-HangzhouUbuchwepheshe be-Bioer) I-MA-6000 Real-Time Quantitative Thermal Cycler (Suzhou Molarray Co., Ltd.) Uhlelo lwe-BioRad CFX96 Real-Time PCR Uhlelo lwe-BioRad CFX Opus 96 lwe-Real-Time PCR |
Ukugeleza Komsebenzi
Inketho 1.
Engeza i-180μL ye-lysozyme buffer ku-precipitate (hlanganisa i-lysozyme ibe ngu-20mg/mL nge-lysozyme diluent), i-pipette ukuze ixube kahle, bese uyicubungula ku-37°C imizuzu engaphezu kwengu-30. Thatha i-1.5mL ye-RNase/DNase-free centrifuge tube, bese ufaka180μL yokulawula okuhle kanye nokulawula okubi ngokulandelana. Engeza10I-μL yokulawula kwangaphakathi kusampula ezohlolwa, ukulawula okuhle, kanye nokulawula okungekuhle ngokulandelana, bese usebenzisa i-Nucleic Acid Extraction noma i-Purification Reagent (YDP302) yi-Tiangen Biotech (Beijing) Co., Ltd. ukuze uthole isampula elandelayo yokukhipha i-DNA, futhi sicela ulandele ngokuqinile imiyalelo yokusebenzisa ngezinyathelo ezithile. Sebenzisa i-DNase/RNase H engenayo i-H2O ye-elution, kanti ivolumu ye-elution enconywayo ingu-100μL.
Inketho 2.
Thatha i-1.5mL yeshubhu ye-centrifuge engena-RNase/DNase, bese ufaka i-200μL yokulawula okuhle kanye nokulawula okubi ngokulandelana.10I-μL yokulawula kwangaphakathi kusampula ezohlolwa, ukulawula okuhle, kanye nokulawula okungekuhle ngokulandelana, bese usebenzisa i-Macro & Micro-Test Viral DNA/RNA Kit (HWTS-3004-32, HWTS-3004-48, HWTS-3004-96) kanye ne-Macro & Micro-Test Automatic Nucleic Acid Extractor (HWTS-3006). Ukukhipha kufanele kwenziwe ngokuhambisana nemiyalelo yokusebenzisa, futhi ivolumu yokususa enconywayo ingu-80μL.
