NEW YORK (GenomeWeb) –Check-Points Health, a Dutch in vitro diagnostic firm that has parlayed its microarray and PCR technologies and expertise into various CE-IVD-marked assays for antibiotic resistance in Gram-negative bacteria, is now looking to move its products into the US market.
To that end, the firm has partnered with a small Chicago-area outfit called Innovative Molecular Diagnostic Systems to begin the process of filing for 510(k) clearance of Check-Direct CPE, its flagship multiplex real-time PCR assay for carbapenem-resistant bacteria, a company official told GenomeWeb this week.
As is the case with many antibiotics, resistance of medically relevant microorganisms to carbapenems is on the rise. According to recent data collected by the European Centre for Disease Prevention and Control, the incidence rate in the EU of carbapenem resistance of Klebsiella pneumoniae — a major culprit in nosocomial infections — nearly doubled from 4.6 percent in 2010 to 8.3 percent in 2013.
This trend is particularly alarming because carbapenems are a major last-line class of antibiotics used to treat healthcare-associated infections. Recognizing this, both the ECDC and US Centers for Disease Control and Prevention have recommended that healthcare facilities deploy more rapid and accurate methods — i.e. molecular — to screen at-risk patients.
Founded in 2002 in Wageningen, the Netherlands, privately held Check-Points has thus far built its business on improving the identification of multi-drug resistance in Gram-negative bacteria, in particular the resistance mechanisms known as carbapenemases, class C cephalosporinases (AmpC), and extended-spectrum beta-lactamases (ESBLs).
Check-Points' primary discovery platform is its proprietary CE-marked microarray-in-a-tube platform designed to simultaneously detect up to 100 specific resistance markers.
Briefly, probes for these markers are placed along with a sample in a specialized microarray tube, and only ligate to their targets if they match exactly to the template – a single-nucleotide mismatch will prevent ligation.
Then, ligated probes are amplified and labeled, and different probes will hybridize to different positions on the in-tube microarray through a dedicated region of the probes. These hybridized amplification products can then be visualized by colorimetric detection using the Check-Points Tube Reader, and dedicated E-Ads software analyzes the presence or absence of carbapenemases, AmpCs, and ESBLs, as well as the resistance genes present.
Although this array technology is highly accurate in terms of detecting antibiotic resistance, its workflow is not ideal for a clinical laboratory. As such, Check-Points has used the platform both in house and in collaboration with partners to identify targets to include in multiplex real-time PCR assays that can be run on existing qPCR platforms, which are available in many hospital settings.
The company's first such assay, Check-Direct CPE, was launched a few years ago with CE marking in the EU, and detects "KPC, VIM, NDM, and OXA-48 including the emerging OXA-181 variant directly from patient rectal swabs," Margareta Carleros, a marketing manager at Check-Points, told GenomeWeb in an email. "In total over 70 carbapenemase variants are covered, belonging to the four groups (KPC, VIM, NDM and OXA-48)."
She added that the assay is also currently validated for all commonly used real-time PCR platforms, such as Thermo Fisher Scientific's ABI 7500, Bio-Rad's CFX96, Roche's LightCycler 480, Qiagen's Rotor-Gene Q, and Becton-Dickinson's BD MAX.
Check-Points is now looking to expand its business overseas, and as part of that process has begun working with US hospitals and academic institutions to validate its array-based discovery platform and potentially develop new real-time PCR tests.
One such collaboration, with the Mayo Clinic, recently yielded data that was presented in November at the Interscience Conference on Antimicrobial Agents and Chemotherapy in Washington, DC. In this study, Mayo researchers used the Check-MDR CT103 kit to genotype 109 well-characterized isolates of Gram-negative bacilli for ESBL, AmpC, and/or carbapenemase genes.
The Check-MDR CT103 assay accurately typed 106 of 109, or 97 percent of Gram-negative isolates. Perhaps more importantly, in 42 of the isolates tested, or 39 percent, the microarray assay enabled the identification of additional resistance genes originally not known to be present.
"Mayo Clinic is one of the leading hospitals in the US and have committed to excellence in the fight against antibiotic resistance," Carleros said. "The researchers at Mayo Clinic quickly saw the potential and decided to evaluate the Check-MDR CT103 and purchase the microarray system."
Carleros did not disclose whether Check-Points is now conducting additional work with Mayo Clinic on this front, but in general, the company is looking to leverage similar partnerships in the US.
"The microarray assays are widely used among international key opinion leaders in this field," she said. "Our network helps us decide which targets are relevant and thereby guide our development of rapid real-time PCR tests. In this manner we've been able to build our expertise in this field while … develop[ing] assays for different clinical needs."
The company has also established a partnership with Innovative Molecular Diagnostic Systems, which is essentially run by a single executive and entrepreneur, Vijai Pasupuleti, who confirmed with GenomeWeb that the partners are now working toward filing for 510(k) clearance of the Check-Direct CPE PCR assay. Neither Pasupuleti nor Carleros disclosed a timeline for this goal. The test is currently available in the US for research use only.
In the meantime, Check-Points has also recently launched a new PCR assay in the EU — Check-MDR CT103 XL, which features "an extended target menu including resistance genes found in Pseudomonas and Acinetobacter," Careleros said.
The company is also working on a number of additional products, she added. "Currently, we are in the process of launching a multiplex real-time PCR assay for extended-spectrum beta-lactamases, Check-Direct ESBL, that can be run in parallel with Check-Direct CPE," she said. "It's currently CE-IVD for BD MAX only, but we are planning to release it for additional real-time PCR systems in the near future."