Carbapenem Nguzogide Gene (KPC/NDM/OXA 48/OXA 23/VIM/IMP)
Aha ngwaahịa
HWTS-OT045 Carbapenem Resistance Gene (KPC/NDM/OXA 48/OXA 23/VIM/IMP) Ngwa nchọpụta (Fluorescence PCR)
Epidemiology
Ọgwụ nje nke Carbapenem bụ ọgwụ nje β-lactam na-ahụkarị nke nwere ụdị nje nje na-agbasa yana yana ọrụ nje bacteria siri ike. N'ihi nkwụsi ike ya na β-lactamase na obere toxicity, ọ ghọọla otu n'ime ọgwụ nje antibacterial kachasị mkpa maka ọgwụgwọ ọrịa nje siri ike. Carbapenems kwụsiri ike na β-lactamases (ESBLs), chromosomes, na cephalosporinases nke plasmid-mediated (AmpC enzymes).
Ọwa
| PCR-Mix 1 | PCR-Mix 2 | |
| FAM | IMP | VIM |
| VIC/HEX | Njikwa ime | Njikwa ime |
| CY5 | NDM | KPC |
| ROX | OXA48
| OXA23 |
Nka nka
| Nchekwa | ≤-18℃ |
| Ndụ nchekwa | Ọnwa iri na abụọ |
| Ụdị ụdị | Sputum, ógbè dị ọcha, swab rectal |
| Ct | ≤36 |
| CV | ≤5.0% |
| LoD | 103CFU/ml |
| Nkọwapụta | a) Ngwa ahụ na-achọpụta nkọwa ndị na-adịghị mma nke ụlọ ọrụ ahaziri ahazi, na nsonaazụ ya na-emezu ihe ndị a chọrọ na ntinye aka kwekọrọ. b) Nsonaazụ nke ule mmeghachi omume cross-reactivity na-egosi na ngwa a enweghị mmeghachi omume na-adịghị mma na ọrịa ndị ọzọ na-eku ume iku ume, dị ka Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, Streptococcus pneumoniae, Neisseria meningitidis, Staphylococcus aureus, Kleb Acinetobacter junii, Acinetobacter haemolyticus, Legionella pneumophila, Escherichia coli, Pseudomonas fluorescens, Candida albicans, Chlamydia pneumoniae, iku ume adenovirus, Enterococcus, ma ọ bụ sample nwere ndị ọzọ ọgwụ na-eguzogide mkpụrụ ndụ ihe nketa, CTX, SHME, wdg. c) Mgbochi nnyonye anya: Mucin, Minocycline, Gentamicin, Clindamycin, Imipenem, Cefoperazone, Meropenem, Ciprofloxacin Hydrochloride, Levofloxacin, Clavulanic acid, Roxithromycin na-ahọrọ maka nnyonye anya ule, na ya pụta na-egosi na n'elu-e kwuru banyere nnyonye anya bekee na-egosi na n'elu-e kwuru okwu interfering bekee na-enweghị ihe mgbochi nke carba PC. NDM, OXA48, OXA23, VIM, na IMP. |
| Ngwa ndị ọdabara | Ngwa Biosystem 7500 ezigbo oge PCR Sistemu Sistemụ PCR Real-Time QuantStudio®5 SLAN-96P Real-Time PCR Systems (Hongshi Medical Technology Co., Ltd.) Sistemụ PCR Real-Time LightCycler®480 Sistemụ nchọpụta PCR nke LineGene 9600 PlusFQD-96A,HangzhouTeknụzụ bioer) MA-6000 Real-Time Quantitative Thermal Cycler (Suzhou Molarray Co., Ltd.) BioRad CFX96 ezigbo oge PCR sistemụ BioRad CFX Opus 96 PCR ezigbo oge |
Usoro ọrụ
Nhọrọ 1.
Reagenti mmịpụta akwadoro: Macro & Micro-Test General DNA/RNA Kit (HWTS-301)9-50, HWTS-3019-32, HWTS-3019-48, HWTS-3019-96) (nke enwere ike iji Macro & Micro-Test Automatic Nucleic Acid Extractor (HWTS-3006C, HWTS-3006B)) site na Jiangsu Macro & Micro-Test Med-Tech Co., Ltd .. Tinye 200μL nke saline nkịtị na thallus precipitate. Usoro ndị na-esote kwesịrị ịgbaso ntuziaka maka mmịpụta, na olu elution akwadoro bụ100μL.
Nhọrọ 2.
Na-atụ aro mmịpụta reagent: Nucleic Acid mmịpụta ma ọ bụ Purification Reagent (YDP302) site Tiangen Biotech (Beijing) Co., Ltd. The mmịpụta kwesịrị ịmalite n'ụzọ siri ike dabere na nzọụkwụ 2 nke ntụziaka maka ojiji (tinye 200μL nke ihe nchekwa GA na thallus precipitate, na ịma jijiji na-kpamkpam ruo mgbe kwụsịtụrụ thallu). Jiri mmiri efu RNase/DNase maka elution, na olu elution a kwadoro bụ 100μL.
Nhọrọ 3.
Reagent mmịpụta akwadoro: Macro & Micro-Test Sample Release Reagent. Ekwesịrị ịsacha sputum sample site n'ịgbakwunye 1mL nke saline nkịtị na thallus precipitate a kpọtụrụ aha n'elu, centrifuged na 13000r / min maka nkeji 5, ma tụfuo ihe dị elu (dobe 10-20µL nke elu). Maka mpaghara dị ọcha na swab rectal, gbakwunye 50μL nke reagent ntọhapụ ozugbo na thallus precipitate a kpọtụrụ aha n'elu, yana usoro ndị na-esote kwesịrị iwepụta ya dịka ntuziaka maka ojiji.
