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 achọrọ nke ntinye aka kwekọrọ. b) Nsonaazụ nke ule mmeghachi omume cross-reactivity na-egosi na ngwa a enweghị mmeghachi omume na-adịghị mma na nje ndị ọzọ na-eku ume iku ume, dị ka Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, Streptococcus pneumoniae, Neisseria meningitidis, Staphylococcus aureus, Kleb junii, Acinetobacter haemolyticus, Legionella pneumophila, Escherichia coli, Pseudomonas fluorescens, Candida albicans, Chlamydia pneumoniae, iku ume adenovirus, Enterococcus, ma ọ bụ samples nwere ndị ọzọ ọgwụ na-eguzogide mkpụrụ ndụ ihe nketa CTX, mecA, SME, SHV. c) Mgbochi nnyonye anya: Mucin, Minocycline, Gentamicin, Clindamycin, Imipenem, Cefoperazone, Meropenem, Ciprofloxacin Hydrochloride, Levofloxacin, Clavulanic acid, Roxithromycin ka ahọpụtara maka nnyonye anya ule, na nsonaazụ na-egosi na n'elu-e kwuru okwu na-egbochi ihe mmeghachi omume enweghị ihe ọ bụla na-egbochi ihe. iji chọpụta mkpụrụ ndụ ihe nketa carbapenem KPC, NDM, OXA48, OXA23, VIM, na IMP. |
| Ngwa ndị ọdabara | Ngwa Biosystem 7500 ezigbo oge PCR Sistemu Sistemụ PCR ezigbo oge QuantStudio5 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 ka thallus na-ebuli elu.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 ntuziaka maka ojiji (tinye 200μL nke ihe nchekwa GA na thallus precipitate). , ma maa jijiji ruo mgbe thallus kwụsịrị kpamkpam).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.
