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 |
Iiparamitha 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 iintsholongwane zokuphefumla, ezifana ne-Klebsiella pneumoniae, i-Acinetobacter baumannii, i-Pseudomonas aeruginosa, i-Streptococcus pneumoniae, i-Neisseria meningitidis, i-Staphylococcus aureus, i-Klexytocasiella ophilus influenza Acinetobacter junii, Acinetobacter haemolyticus, Legionella pneumophila, Escherichia coli, Pseudomonas fluorescens, Candida albicans, Chlamydia pneumoniae, Respiratory adenovirus, Enterococcus, okanye iisampulu eziqulethe ezinye iigenes ezinganyangekiyo ngamachiza, TMEM, mecSHA etc. 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 ikhethwa kuvavanyo lokuphazamiseka, kwaye iziphumo zibonisa ukuba izinto eziphazamisayo ezikhankanywe ngasentla azikho ukuchasana kwe-carpens ye-KDM, i-KDM i-interfering reaction, i-KDM, 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 kwi-thallus precipitate. Amanyathelo alandelayo kufuneka alandele imiyalelo yokutsalwa, kunye nomthamo we-lution ocetyiswayo100μL.
Ukhetho lwesi-2.
I-reagent yokutsalwa ecetyiswayo: I-Nucleic Acid Extraction okanye i-Purification Reagent (YDP302) yi-Tiangen Biotech (Beijing) Co., Ltd. Ukukhutshwa kufuneka kuqalwe ngokungqongqo ngokuhambelana nenyathelo lesi-2 lomyalelo wokusetyenziswa (yongeza i-200μL ye-buffer GA kwi-thallus precipitate, kwaye ushukume de i-thallus iphelele). 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.
