I-Clostridium difficile toxin A/B gene (C.diff)
Igama lemveliso
Ikhithi yokuFumanisa i-HWTS-OT031A Nucleic Acid yeClostridium difficile toxin A/B gene (C.diff) (Fluorescence PCR)
Isatifikethi
CE
I-Epidemiology
IClostridium difficile (CD), i-gram-positive anaerobic sporogenic Clostridium difficile, yenye yezona pathogens ziphambili ezibangela usulelo lwamathumbu e-nosocomial. Ngokwezonyango, malunga ne-15% ~ 25% yesifo sorhudo esinxulumene neentsholongwane, i-50% ~ 75% yesifo se-antimicrobial-related colitis kunye ne-95% ~ 100% ye-pseudomembranous enteritis zibangelwa yi-Clostridium difficile infection (CDI). IClostridium difficile yipathogen eneemeko ezithile, kuquka iintlobo ze-toxigenic kunye neentlobo ezingezizo ze-toxigenic.
Itshaneli
| I-FAM | i-tcdAijini |
| I-ROX | i-tcdBijini |
| I-VIC/HEX | Ulawulo lwangaphakathi |
Iiparameters zobugcisa
| Indawo yokugcina izinto | ≤-18℃ |
| Beka ubomi kwishelufa | Iinyanga ezili-12 |
| Uhlobo lweSibonelelo | isitulo |
| Tt | ≤38 |
| CV | ≤5.0% |
| I-LoD | 200CFU/mL |
| Ukuchaneka | Sebenzisa le khithi ukubona ezinye izifo zamathumbu ezifana ne-Escherichia coli, i-Staphylococcus aureus, i-Shigella, i-Salmonella, i-Vibrio parahaemolyticus, i-Group B Streptococcus, i-Clostridium difficile non-pathogenic strains, i-Adenovirus, i-rotavirus, i-norovirus, i-influenza A virus, i-influenza B virus kunye ne-human genomic DNA, zonke iziphumo azilunganga. |
| Izixhobo Ezisebenzayo | Iinkqubo ze-PCR ze-Applied Biosystems 7500 Real-Time Iinkqubo ze-PCR eziSebenzisiweyo ze-7500 ezikhawulezayo zeXesha langempela I-QuantStudio®Iinkqubo ezi-5 ze-PCR zeXesha langempela Iinkqubo ze-PCR ze-SLAN-96P zeXesha langempela (iHongshi Medical Technology Co., Ltd.) I-LightCycler®Inkqubo ye-PCR yeXesha langempela ye-480 Inkqubo yokuHlola i-LineGene 9600 Plus yeXesha langempela (I-FQD-96A，IHangzhouItekhnoloji yeBioer) I-MA-6000 Real-Time Quantitative Thermal Cycler (Suzhou Molarray Co., Ltd.) Inkqubo ye-PCR ye-BioRad CFX96 yeXesha langempela Inkqubo ye-PCR ye-BioRad CFX Opus 96 yeXesha langempela |
Ukuhamba komsebenzi
Ukhetho 1.
Yongeza i-180μL ye-lysozyme buffer kwi-precipitate (xuba i-lysozyme ibe yi-20mg/mL nge-lysozyme diluent), i-pipette ukuze ixutywe kakuhle, kwaye uyixube kwi-37°C ngaphezulu kwemizuzu engama-30. Thatha i-1.5mL yetyhubhu ye-centrifuge engena-RNase/DNase, uze wongeze180μL yolawulo oluhle kunye nolawulo olubi ngokulandelelana. Yongeza10I-μL yolawulo lwangaphakathi kwisampulu eza kuvavanywa, ulawulo oluqinisekileyo, kunye nolawulo olungalunganga ngokulandelelana, kwaye usebenzise i-Nucleic Acid Extraction okanye i-Purification Reagent (YDP302) yiTiangen Biotech (Beijing) Co., Ltd. kwisampulu elandelayo yokukhupha i-DNA, kwaye nceda ulandele ngokungqongqo imiyalelo yokusetyenziswa kwamanyathelo athile. Sebenzisa i-DNase/RNase free H2U-O kwi-elution, kwaye umthamo we-elution ocetyiswayo yi-100μL.
Ukhetho 2.
Thatha i-1.5mL yetyhubhu ye-centrifuge engena-RNase/DNase, uze wongeze i-200μL yokulawula okulungileyo kunye nokulawula okubi ngokulandelelana.10I-μL yolawulo lwangaphakathi kwisampulu eza kuvavanywa, ulawulo oluqinisekileyo, kunye nolawulo olungalunganga ngokulandelelana, kwaye usebenzise iMacro & Micro-Test Viral DNA/RNA Kit (HWTS-3004-32, HWTS-3004-48, HWTS-3004-96) kunye neMacro & Micro-Test Automatic Nucleic Acid Extractor (HWTS-3006). Ukukhupha kufuneka kwenziwe ngokuhambelana ngqo nemiyalelo yokusetyenziswa, kwaye umthamo wokucoca ocetyiswayo yi-80μL.
