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Klebsiella Pneumoniae, Acinetobacter Baumannii and Pseudomonas Aeruginosa and Drug Resistance Genes (KPC, NDM, OXA48 and IMP) Multiplex Detection Kit (Fluorescence PCR)

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Klebsiella Pneumoniae, Acinetobacter Baumannii and Pseudomonas Aeruginosa and Drug Resistance Genes (KPC, NDM, OXA48 and IMP) Multiplex Detection Kit (Fluorescence PCR)

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Introduction

[Product Name] Klebsiella Pneumoniae, Acinetobacter Baumannii and Pseudomonas Aeruginosa and Drug Resistance Genes (KPC, NDM, OXA48 and IMP) Multiplex Detection Kit (Fluorescence PCR)
[Packaging Size] 20 tests/kit, 50 tests/kit

[Intended Use]

This kit is used for in vitro qualitative detection of Klebsiella pneumoniae (KPN), Acinetobacter baumannii (Aba), Pseudomonas aeruginosa (PA) and four carbapenem resistance genes (which include KPC, NDM, OXA48 and IMP) in human sputum samples, to provide the basis of guidance of clinical diagnosis, treatment and medication for patients with suspected bacterial infection.

Klebsiella pneumoniae is a common clinical opportunistic pathogen and one of the important pathogenic bacteria causing nosocomial infections. When the body's resistance is reduced, the bacteria enter the lungs from the respiratory tract, causing infection in multiple parts of the body, and the early use of antibiotics is the key to cure [1]. Klebsiella pneumoniae is resistant to commonly antibacterial drugs (such as aminoglycosides, fluoroquinolones, fosfomycins, etc.), as well as carbapenems that known as the ' last line of defense ' of antibiotics, with an average of 10.9% nationwide. The available therapeutic drugs are very limited (such as polymyxin and tigecycline), resulting in a high fatality rate of patients and a serious threat to the life and health of patients [2].

The most common site of Acinetobacter baumannii infection is the lungs, which is an important pathogen for Hospital acquired pneumonia (HAP), especially Ventilator associated pneumonia (VAP). It is often accompanied by other bacterial and fungal infections, with characteristics of high morbidity rate and high mortality rate. Acinetobacter baumannii has strong ability of acquired resistance and monoclonal spread, and has become one of the most important pathogens of nosocomial infection. The National Drug Resistance Surveillance Network shows that the national average resistance rate of Acinetobacter baumannii to carbapenems is 53.7%.

Pseudomonas aeruginosa is the most common non-fermentative gram-negative bacilli in clinical practice, and is an important opportunistic pathogen for hospital-acquired infection, with the characteristics of easy colonization, easy variation and multi-drug resistance. The lower respiratory tract is the most common site of nosocomial bacterial infection. The types of infections mainly include bronchiectasis complicated with infection, Chronic obstructive pulmonary disease (referred to as COPD) complicated with infection and pneumonia, etc. Lower respiratory tract infections caused by multi-drug resistant PA have a high mortality rate and are difficult to treat [3]. The Carbapenems resistance Pseudomonas aeruginosa (CRPA) remains high. The global bacterial resistance monitoring data shows that CRPA ranks among the top pathogens of HAP. At the same time, the resistance rate to commonly used antibiotics is increasing year by year, with an average of 18.3% nationwide, which has caused great trouble to clinical treatment.

Carbapenems are atypical β-lactam antibiotics with the broadest antibacterial spectrum and the strongest antibacterial activity. Because of the stability to β-lactamase and low toxicity, Carbapenems have become one of the main antibacterial drugs for treating serious bacterial infections. Carbapenems are highly stable to plasmid-mediated Extended-spectrum β-lactamases (ESBLs), chromosome and plasmid-mediated cephalosporinase (AmpC enzymes). The main cause of producing drug resistance is carbapenemase. Carbapenem enzymes can be divided into three categories class A, class B and class D according to Ambler classification. Among them, class A and class D are serine enzymes, and class B are metalloenzymes. The most common class A enzymes are Klebsiella pneumoniae carbapenemases (KPC) [4], and class B enzymes are New Delhimetalloenzymes (NDM) and class D enzymes are Oxacillinases (OXA) [5].

This kit provides an auxiliary basis for the diagnosis of various common bacterial infections and Carbapenems resistance drugs. The test results are for clinical reference only, and the final diagnosis must be closely combined with other clinical indicators for comprehensive analysis.

[Test Principles]

This kit uses fluorescent PCR method to design primer probes of fluorescence detection for the conservative sequences of KPN, Aba and PA and carbapenem resistance genes KPC, NDM, OXA48 and IMP, and the probes are labeled with fluorescent groups at the 5’ end and labeled with quencher groups at the 3’ end. During the detection and amplification process, specific primers and probes bind to their respective target sequences separately, and the formation of PCR products and the accumulation of fluorescent signals are completely synchronized by the DNA polymerase activity of Taq enzyme and 5'-3' exonuclease activity, realizing the qualitative detection of KPN, Aba, PA and drug resistance genes KPC, NDM, OXA48 and IMP in the samples. At the same time, endogenous internal reference quality control is introduced to label VIC/HEX fluorescent groups to avoid false negative in detection.

[Storage Conditions and Shelf-life]

Storage condition: This kit should be stored below -18°C away from light. The shelf life is 12 months. Avoid the repeated freezing and thawing (no more than 4 cycles). After opening, if it is stored at 2~8°C, please use within 1 week.
Shipping conditions: The kit is stable for 5 days in a shipping box containing dry ice.

See the packaging label for the production date, production batch number and expiry date.

[Applicable Instruments]

Applied Biosystems 7500 Real-Time PCR Systems, Applied Biosystems 7500 Fast Real-Time PCR Systems, QuantStudio®5 Real-Time PCR Systems, SLAN-96P Real-Time PCR Systems (Shanghai Hongshi Medical Technology Co., Ltd.), LightCycler®480 Real-Time PCR system, LineGene 9600 Plus Real-Time PCR Detection System (FQD-96A, Bioer technology), MA-6000 Real-Time Quantitative Thermal Cycler (Suzhou Molarray Co., Ltd.), BioRad CFX96 Real-Time PCR System, and BioRad CFX Opus 96 Real-Time PCR System.

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