I-Carbapenem Resistance Gene (KPC/NDM/OXA 48/OXA 23/VIM/IMP)
Igama lemveliso
I-HWTS-OT045 Carbapenem Resistance Gene (KPC/NDM/OXA 48/OXA 23/VIM/IMP) Ikhithi yokuFumana (i-Fluorescence PCR)
I-Epidemiology
I-Carbapenem antibiotics i-atypical β-lactam antibiotics kunye ne-antibacterial spectrum ebanzi kunye nomsebenzi onamandla wokulwa ne-antibacterial.Ngenxa yokuzinza kwayo kwi-β-lactamase kunye netyhefu ephantsi, iye yaba yenye yezona ziyobisi zibalulekileyo kunyango losulelo olubi lwebhaktiriya.I-Carbapenems izinzile kakhulu kwi-plasmid-mediated extended-spectrum β-lactamases (ESBLs), i-chromosomes, kunye ne-plasmid-mediated cephalosporinases (i-AmpC enzymes).
Umjelo
| I-PCR-Mix 1 | PCR-Mix 2 | |
| FAM | IMP | VIM |
| VIC/HEX | Ulawulo lwangaphakathi | Ulawulo lwangaphakathi |
| CY5 | NDM | I-KPC |
| ROX | OXA48
| OXA23 |
IiParameters zobuGcisa
| Ugcino | ≤-18℃ |
| Beka ubomi kwishelufa | Iinyanga ezili-12 |
| Uhlobo loMfanekiso | Isikhohlela, iikholoni ezicocekileyo, i-rectal swab |
| Ct | ≤36 |
| CV | ≤5.0% |
| LoD | 103CFU/mL |
| Ukuchaza ngokuthe ngqo | a) Ikhithi ibona izalathisi ezingalunganga zenkampani, kwaye iziphumo ziyahlangabezana neemfuno zeereferensi ezihambelanayo. b) Iziphumo zovavanyo lwe-cross-reactivity zibonisa ukuba le kit ayinayo impendulo emnqamlezweni kunye nezinye izifo zokuphefumla, ezifana ne-Klebsiella pneumoniae, i-Acinetobacter baumannii, i-Pseudomonas aeruginosa, i-Streptococcus pneumoniae, i-Neisseria meningitidis, i-Staphylococcus aureus, i-Klexytobaccus aureus, i-Klexytobacinela, i-Klexytobacinela, i-Klexytobacinela, i-Klexytobacinela junii, Acinetobacter haemolyticus, Legionella pneumophila, Escherichia coli, Pseudomonas fluorescens, Candida albicans, Chlamydia pneumoniae, Respiratory adenovirus, Enterococcus, okanye iisampuli eziqulethe ezinye iintlobo ze-CTX, mecVV, TEM, njl njl. c) I-Anti-interference: i-Mucin, i-Minocycline, i-Gentamicin, i-Clindamycin, i-Imipenem, i-Cefoperazone, i-Meropenem, i-Ciprofloxacin Hydrochloride, i-Levofloxacin, i-Clavulanic acid, i-Roxithromycin ikhethiweyo kuvavanyo lokuphazamiseka, kwaye iziphumo zibonisa ukuba izinto eziphazamisayo ezikhankanywe ngasentla azikho ukusabela okuphazamisayo. ukufumanisa i-carbapenem resistance genenes KPC, NDM, OXA48, OXA23, VIM, kunye ne-IMP. |
| Izixhobo Ezisebenzayo | Iinkqubo ze-Biosystems ezisetyenzisiweyo 7500 Iinkqubo zePCR zeXesha langempela I-QuantStudio®5 Iinkqubo zePCR zeXesha langempela SLAN-96P Iinkqubo zePCR zeXesha elililo (Hongshi Medical Technology Co., Ltd.) Inkqubo yePCR yeLightCycler®480 yeXesha langempela I-LineGene 9600 Plus ye-Real-Time PCR yokuFumana iNkqubo (FQD-96A,HangzhouItekhnoloji yeBioer) I-MA-6000 yeXesha lokwenyani iBhayisekile yoMyinge weThermal (iSuzhou Molarray Co., Ltd.) I-BioRad CFX96 Inkqubo yePCR yexesha lokwenyani BioRad CFX Opus 96 Real-Time PCR System |
Ukuhamba komsebenzi
Ukhetho loku-1.
I-reagent ecetyiswayo yokutsalwa: iMacro & Micro-Test General DNA/RNA Kit (HWTS-3019-50, HWTS-3019-32, HWTS-3019-48, HWTS-3019-96) (enokuthi isetyenziswe kunye ne-Macro & Micro-Test Automatic Nucleic Acid Extractor (HWTS-3006C, HWTS-3006B)) yi-Jiangsu Macro & Micro-Test Med-Tech Co., Ltd.. Yongeza i-200μL ye-saline eqhelekileyo ukuya imvula yethallus.Amanyathelo alandelayo kufuneka alandele imiyalelo yokutsalwa, kunye nomthamo we-lution ocetyiswayo100μL.
Ukhetho lwesi-2.
Ukukhutshwa kwe-reagent ecetyiswayo: I-Nucleic Acid Extraction okanye i-Reagent yokucoca (YDP302) yi-Tiangen Biotech (Beijing) Co., Ltd. Utsalo kufuneka luqalwe ngokungqongqo ngokuhambelana nenyathelo lesi-2 lomyalelo wokusetyenziswa (yongeza i-200μL ye-buffer GA kwimvula yethallus. , kwaye ungcangcazele de i-thallus imiswe ngokupheleleyo).Sebenzisa iRNase/DNase amanzi asimahla elution, kunye nomthamo we-lution onconywayo yi-100μL.
Ukhetho lwesi-3.
I-reagent yokutsalwa ecetyiswayo: I-Macro kunye ne-Micro-Test yeSampule yokuKhupha iReagent.Isampulu yesikhohlela kufuneka ihlanjwe ngokugalela i-1mL ye-saline eqhelekileyo kwi-thallus precipitate enyangwayo ekhankanywe ngasentla, ifakwe phakathi kwi-13000r/min imizuzu emi-5, kwaye i-supernatant iyalahlwa (gcina i-10-20µL ye-supernatant).Kwikholoni ecocekileyo kunye ne-rectal swab, yongeza i-50μL ye-reagent yokukhulula isampuli ngokuthe ngqo kwi-thallus precipitate ekhankanywe ngasentla, kwaye amanyathelo alandelayo kufuneka akhutshwe ngokomyalelo wokusetyenziswa.
