AffiPCR® MDR KPC/OXA Real Time PCR

Target Disease Type: Bacterial drug resistance

Specific Application: Drug Resistance

Storage and Shipping : 4 weeks

MDR KPC/OXA RT-PCR (CE) C2-100FRT DataSheet

INTRODUCTION

Nosocomial respiratory tract infections are major cause of excessive morbidity and mortality. Patients with serious underlying diseases have an especially high risk of acquiring these infections and that risk is magnified by exposure to respiratory therapy.

Beta-lactams remain a cornerstone for antimicrobial chemotherapy of a large number of bacterial infections, but their efficacy has been increasingly thwarted by dissemination of acquired resistance determinants among pathogenic bacteria. The exposure of bacterial strains to a multitude of β-lactams has induced a dynamic and continuous production and mutation of β-lactamase in many bacteria, expanding their activity even against later generation cephalosporins 5 and carbapenems by the production of extended-spectrum beta-lactamase (ESBL) and metallo-beta-lactamase (MBL) respectively. Since the genes that code for the production of ESBL are often linked to other resistance genes causing extended spectrum of drug resistance, this will result into fewer therapeutic alternatives.

ESBLs with hydrolytic activity against carbapenems are classified in three groups:

1. Class A β-lactamases - Klebsiella pneumoniae carbapenemase (KPC)
2. Class B - metallo-beta-lactamase (MBL) which includes New Delhi metallo-β-lactamase (NDM), Verona integron-encoded-metallo-β-lactamase (VIM) and imipenemase-metallo-β-lactamase (IMP)
3. Class D - OXA-carbapenemases.

Main ESBL-producing pathogens are:

• Enterobacteriaceae
• coli
• Pneumoniae
• Oxytoca
• mirabilis
• Enterobacter
• Salmonella
• Non-fermentative Gram-negative
• baumannii
• aeruginosa

Methods of detection of ESBLs:

• Phenotypic methods (antibiotic susceptibility)
• Used routinely in clinical laboratories
• The accuracy of semiautomated microbiology systems is not optimal
• Genotypic methods (PCR-based amplification)
• Used in reference laboratories
• Discriminate between specific types of ESBLs
• Need shorter time to detection (culture not required)
• Have ability to detect low level resistance

INTENDED USE

MDR KPC/OXA Real-TM PCR kit is an in vitro nucleic acid amplification test for detection and differentiation of MDR genes using real-time hybridization-fluorescence detection of amplified products.

PRINCIPLE OF ASSAY

The detection of MDR genes includes DNA isolation from biological materials and real-time PCR amplification of DNA. MDR genes detection by the polymerase chain reaction (PCR) is based on the amplification of genome specific region using specific primers. In real-time PCR, the amplified product is detected using fluorescent dyes. These dyes are linked to oligonucleotide probes which bind specifically to the amplified product. The real-time monitoring of fluorescence intensities during the real-time PCR allows detection of the amplified product without re-opening the reaction tubes after the PCR run. MDR KPC/OXA Real-TM PCR kit uses “hot-start”, which greatly reduces the frequency of nonspecifically primed reactions.

• The carbapenemase group KPC is detected in the FAM/Green channel.
• The carbapenemase group OXA-48-similar (OXA-48 and OXA-162) is detected in the JOE/HEX/Yellow channel
• The Internal Control (IC) is detected in the Rox/Texas Red/Orange channel.