Gene yo kurwanya Carbapenem (KPC/NDM/OXA 48/OXA 23/VIM/IMP)
Izina ry'igicuruzwa
HWTS-OT045 Carbapenem Resistance Gene (KPC/NDM/OXA 48/OXA 23/VIM/IMP) Imashini yo Gusuzuma (Fluorescence PCR)
Ubumenyi bw'indwara
Antibiyotike za Carbapenem ni antibiyotike zidasanzwe za β-lactam zifite imiterere yagutse ya antibiyotike kandi zifite ubushobozi bukomeye bwo kurwanya antibiyotike. Kubera ko zihamye kuri β-lactamase kandi zikaba nkeya, zabaye imwe mu miti ikomeye ya antibiyotike mu kuvura indwara zikomeye za bagiteri. Carbapenems ihamye cyane kuri β-lactamase (ESBLs), chromosomes, na cephalosporinases (AmpC enzymes) za plasmid.
Umuyoboro
| PCR-Uruvange rwa 1 | PCR-Mix 2 | |
| FAM | IMP | VIM |
| VIC/HEX | Igenzura ry'imbere | Igenzura ry'imbere |
| CY5 | NDM | KPC |
| ROX | OXA48
| OXA23 |
Ibipimo bya tekiniki
| Ububiko | ≤-18℃ |
| Igihe cyo kumara | Amezi 12 |
| Ubwoko bw'ingero | Amashoka, amakorosi meza, agapfukamunwa k'umurambo |
| Ct | ≤36 |
| CV | ≤5.0% |
| LoD | 103CFU/mL |
| Ubwihariye | a) Iyi kit isuzuma ibimenyetso bibi by’ikigo, kandi ibisubizo byujuje ibisabwa n’ibimenyetso bihuye nabyo. b) Ibisubizo by'ikizamini cya "cross-reactivity" bigaragaza ko iyi kit nta kibazo ifite ku zindi ndwara zitera indwara z'ubuhumekero, nka Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, Streptococcus pneumoniae, Neisseria meningitidis, Staphylococcus aureus, Klebsiella oxytoca, Haemophilus influenzae, Acinetobacter junii, Acinetobacter haemolyticus, Legionella pneumophila, Escherichia coli, Pseudomonas fluorescens, Candida albicans, Chlamydia pneumoniae, Respiratory adenovirus, Enterococcus, cyangwa ingero zirimo izindi gene zidashobora imiti CTX, mecA, SME, SHV, TEM, nibindi. c) Kurwanya ingaruka mbi: Mucin, Minocycline, Gentamicin, Clindamycin, Imipenem, Cefoperazone, Meropenem, Ciprofloxacin Hydrochloride, Levofloxacin, aside Clavulanic, Roxithromycin ni byo byatoranijwe kugira ngo bipimwe ingaruka mbi, kandi ibisubizo bigaragaza ko ibintu bivangura byavuzwe haruguru nta ngaruka mbi bigira mu gutahura jeni zirwanya carbapenem KPC, NDM, OXA48, OXA23, VIM, na IMP. |
| Ibikoresho Bikoreshwa | Sisitemu za PCR zikoreshwa mu gihe nyacyo 7500 Sisitemu za PCR za QuantStudio®5 mu gihe nyacyo Sisitemu za PCR za SLAN-96P mu gihe nyacyo (Hongshi Medical Technology Co., Ltd.) Sisitemu ya LightCycler®480 Real-Time PCR Sisitemu yo Gusuzuma PCR ya LineGene 9600 Plus mu gihe nyacyo (FQD-96A，HangzhouIkoranabuhanga rya Bioer) MA-6000 Real-Time Quantitative Thermocycler (Suzhou Molarray Co., Ltd.) Sisitemu ya BioRad CFX96 ya PCR mu gihe nyacyo Sisitemu ya BioRad CFX Opus 96 ya PCR mu gihe nyacyo |
Urugendo rw'akazi
Amahitamo ya 1.
Reagent isabwa gukurwamo: Macro & Micro-Test General DNA/RNA Kit (HWTS-301)9-50, HWTS-3019-32, HWTS-3019-48, HWTS-3019-96) (ishobora gukoreshwa hamwe na Macro & Micro-Test Automatic Nucleic Acid Extractor (HWTS-3006C, HWTS-3006B)) na Jiangsu Macro & Micro-Test Med-Tech Co., Ltd.. Shyiramo 200μL y'umunyu usanzwe kuri thallus precipitate. Intambwe zikurikiraho zigomba gukurikiza amabwiriza yo gukuramo, kandi ingano ya elution isabwa ni100μL.
Amahitamo ya 2.
Reagent isabwa gukurwamo: Reagent yo gukuramo aside ya Nucleic cyangwa Purification (YDP302) ya Tiangen Biotech (Beijing) Co., Ltd. Gukuramo bigomba gutangizwa hakurikijwe neza intambwe ya 2 y'amabwiriza yo gukoresha (ongeramo 200μL ya buffer GA kuri thallus precipitate, hanyuma uzunguze kugeza thallus ihagaritswe burundu). Koresha amazi ya RNase/DNase adakoresha kugira ngo uhuze, kandi ingano ya elution igenwa ni 100μL.
Amahitamo ya 3.
Reagent isabwa gukurwamo: Reagent yo gukuramo icyitegererezo cya Macro na Micro-Test. Icyitegererezo cy'amatembabuzi kigomba kozwa wongeyeho 1mL y'umunyu usanzwe kuri thallus precipitate yavuzwe haruguru, ushyirwa kuri centrifuge kuri 13000r/min mu gihe cy'iminota 5, hanyuma supernatant ikajugunywa (bika 10-20µL ya supernatant). Ku ifu y'uruhu n'iy'umurambo, ongeramo 50μL ya reagent yo gukuramo icyitegererezo ku thallus precipitate yavuzwe haruguru, hanyuma intambwe zikurikira zigomba gukurwamo hakurikijwe amabwiriza yo kuyikoresha.
