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KPC/IMP/NDM/VIM/OXA-48 Combo Test Kit (LFIA)
● Instrumentation: not required
● Detection time: 20-25 minutes
● Storage: Stored away from direct sunlight at 2℃ to 30℃
● Shelf life: 24 months
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Intended Use
KPC/IMP/NDM/VIM/OXA-48 Combo Test Kit (LFIA) is a colloidal gold immunochromatography for the rapid qualitative detection of KPC, IMP, NDM, VIM and OXA-48 carbapenemases in bacterial samples obtained after culture.
Summary
Antimicrobial resistance (AMR) poses a serious global threat of growing concern to human, animal, and environment health. This is due to the emergence, spread, and persistence of multidrug-resistant (MDR) bacteria or “superbugs.”
Several fields of modern medicine depend on the availability of effective antibiotic drugs; chemotherapy for cancer treatment, organ transplantation, hip replacement surgery, intensive care for pre-term newborns and many other activities could not be performed without effective antibiotics. In fact, infections caused by multidrug-resistant bacterial strains are among the main factors influencing morbidity and mortality in patients undergoing these procedures.
Enterobacterales are conditionally pathogenic bacteria that cause serious hospital-acquired infections. The spread of carbapenemase-producing Enterobacterales (CPE) has become a major global public health threat. Carbapenems have traditionally been used to treat infections caused by broad-spectrum beta-lactamase-producing Escherichia coli and Klebsiella pneumoniae and are still considered antibiotics to be used as a last resor. Carbapenemase-producing enzymes in these bacteria, which are capable of hydrolyzing all carbapenems, cephalosporins, and beta-lactams are the main cause of resistance to carbapenem antibiotics. Most carbapenemase genes are located on the metastable genetic elements, such as plasmids and integrons; thus, carbapenem resistance is easily transferred horizontally leading to rapid spread wordwide.
At present, the common carbapenemases mainly include KPC type, IMP type, NDM type, VIM type and OXA-48 type.
Test Principle
KPC/IMP/NDM/VIM/OXA-48 Combo Test Kit (LFIA) detects the five most common carbapenemase families (KPC, IMP, NDM, VIM and OXA-48) with colloidal gold immunochromatography using a double antibody sandwich assay.
When the appropriate amount of test samples treated with dilution buffer is added to the sample well of the test cassette, the sample will move forward along the test strip by capillary action. If the sample contains KPC, IMP, NDM, VIM and OXA-48 carbapenemases and the concentration is higher than the limit of detection, the antigen will form immune complexes with corresponding carbapenemase antibody labeled with colloidal gold respectively, which are captured by K line, I line, N line, V line and O line. If test sample contains KPC carbapenemase, forming a red K line, indicating a positive result for KPC carbapenemase. If test sample contains IMP carbapenemase, forming a red I line, indicating a positive result for IMP carbapenemase. If test sample contains NDM carbapenemase, forming a red N line, indicating a positive result for NDM carbapenemase. If test sample contains VIM carbapenemase, forming a red V line, indicating a positive result for VIM carbapenemase. If test sample contains OXA-48 carbapenemase, forming a red O line, indicating a positive result for OXA-48 carbapenemase.
Additionally, the test strip also contains a control line (C line). The C line should be formed to indicate that the sample has been transported properly through the membrane regardless of whether sample contains carbapenemases or not. If the C line does not appear, it indicates that the test result is invalid and the sample need to retest.
Test Procedure
1.Scrape one loop of bacteria with a 1ul inoculating loop, and the scraped bacteria should be full of inoculation loop hole as far as possible.
2.Dip the loop in the bottom of the tube containing the dilution buffer. Shake the inoculating loop in order to wash off the bacteria in the tube as much as possible. lf the bacteria are sticky and difficult to wash off, a vortex mixer can be used to fully wash the bacteria off the inoculation loop.
Repeat the above steps to scrape 2 loops of bacteria and wash them off in the dilution buffer.
3. Remove the inoculating loop, cover the tube with the dropper and mix well on the vortex mixer.
Attention: The normalization of sampling and the adequacy of processing samples into dilution buffer directly affect the test results.
4. Open the aluminum foil pouch, take out the test cassette and lay it on a clean flat surface. Add 4 drops (approximately 100 μL) processed sample extract into the sample well. The results should be observed within 15-20 minutes. Result observed after 20 minutes is invalid.
Remark: Additional required but not provided equipment: Timer, inoculating loop, culture dish and vortex mixer.
Test Method Limitations
1. This product is only for qualitative testing and the specific concentration of each indicator must be measured using other quantitative methodologies.
2. This test is used to qualitatively detect KPC, IMP, NDM, VIM and OXA-48 carbapenemases in bacterial samples. A positive or negative result does not rule out the existence of other antibiotic resistance mechanisms.
3. Improper handling and use will affect the test results of the samples.
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