NEW YORK ─ Bremen, Germany-based Molzym, a developer of PCR-based diagnostic test kits for European markets, is planning to enter the US in a few years with an automated, multiplex sepsis test that detects a broad range of pathogens directly from patient samples within five hours.
Molzym is developing the instrument and panel in collaboration with Munich-based Fraunhofer Institute as part of a €2 million ($2.4 million) project funded by the European Union's European Regional Development Fund and the federal state of Brandenburg, Germany.
To identify microbes that cause infections, labs generally need to conduct a lengthy process for culturing cells. It can take eight hours or more before a laboratory can run one test to identify the pathogen and run another, an antimicrobial susceptibility test, that enables a clinician to decide whether to prescribe an antibiotic and which one to prescribe.
As part of a testing strategy to identify infectious disease pathogens directly from samples and speed the time to identification, Molzym and Fraunhofer are developing a platform, called AutoSepT, that removes unwanted host DNA and incorporates specially designed PCR primers to detect sepsis and other infectious microbes.
"In patient samples, as you can imagine, there's an overload of host DNA that can be a strong inhibitor of test performance," Michael Lustig, Molzym's COO, said in an interview. "We've developed a process to significantly reduce host DNA background."
Molzym uses a special chemical ─ a high chaotropic buffer ─ to lyse host cells to remove unwanted DNA, Lustig said.
Its PCR primers target 16s rRNA to detect and identify bacterial pathogens and 18s rRNA to detect and identify fungal pathogens, and its technology further leverages contamination-free PCR reagents, buffers, and enzymes that are critical to conducting broad-range pathogen analyses, he added.
Molzym and Fraunhofer are focusing first on integrating bacterial targets in the sepsis panel, but they plan to integrate fungal targets later.
Lustig said that the collaborators have already developed a capability for automated DNA extraction, but now they need to integrate the analysis and interpretation of results into a fully automated system.
For the project with Fraunhofer, Molzym has opened a development facility in Potsdam-Golm, Germany, where it will continue developing the platform and prepare it for future clinical trials and regulatory approvals worldwide. The firm has plans to launch the test in Europe, the US, Asia, and the Middle East. Future clinical trials would need to be robust enough to satisfy stringent, upcoming IVDR regulations in Europe, and US Food and Drug Administration 510(k) clearance in the US, which could come around 2023, Lustig said.
Molzym has approximately 20 employees including doctors, researchers, technicians, and laboratory specialists working on molecular microbiology research and routine microbial pathogen diagnostic test systems. The firm has been in business since 2003 with sales mainly in Europe, where it markets its tests through distribution agreements.
In December 2019, Miami-based Gradian Diagnostics, a holding company focused on developing medical and diagnostics technologies, acquired Molzym for an undisclosed fee. Gradian Dx brought financial and managerial knowledge and experience to help Molzym grow its business and sales activities, Lustig said, adding that the parent company is supporting its fundraising and the automation of test production.
Unbiased identification
Molzym has been working on technology that avoids the use of cell culturing since it introduced its first test, SepsiTest UMD, for which it obtained CE marking in 2008. The test detects and identifies a broad range of gram-positive bacteria, gram-negative bacteria, and fungi that lead to infectious disease in body fluids, tissues, and swabs — and provides results within a day, according to the firm.
Instead of cell culturing, it uses a manual process to selectively isolate microbial DNA and then real-time PCR to amplify the DNA targets prior to Sanger sequencing for pathogen detection and identification.
Among Molzym's customers, Copenhagen University Hospital implemented its UMD assay in 2015, and the test "has shown to be extremely valuable for the identification of pathogens in culture-negative samples such as infective endocarditis, bone and joint infections, otitis media, and meningitis, or samples with initial suspicion of a low number of bacteria," said Thomas Bjarnsholt, a professor in the Department of Immunology and Microbiology at the University of Copenhagen.
Lustig said that at the time of its introduction, the company envisioned the test would achieve the greatest traction for sepsis testing, but that didn't pan out. Use of Sanger sequencing contributed to the broad detection and identification of bacterial and fungal pathogens, but required a full day or night to obtain results — too much time to be practical for clinicians looking to quickly diagnose sepsis, Lustig said.
As a result, Molzym has continued to develop its technology for sepsis and other infectious diseases, and in 2017 it obtained CE marking for a new diagnostic kit, Micro-Dx, which uses the same technology as SeptiTest but is integrated with an automated instrument called SelectNA plus that enables automated DNA preparation.
However, Micro-Dx still relies on Sanger sequencing for final results, and the new test being developed with the Fraunhofer Institute should align better with the needs of clinicians treating sepsis patients, Lustig said.
Within five hours, the multiplex panel would detect and identify all microbes that cause about 90 percent of sepsis cases, enabling a clinician to make treatment decisions at that time. Samples are still processed by Sanger sequencing to detect and identify the additional 10 percent or so of pathogens that cause sepsis as well as rare pathogens, Lustig said.
Such an approach is likely to be far more acceptable than waiting overnight for all results, Lustig said. Further, the platform is being developed to operate with a range of samples including blood, urine, nasal swabs, tissue, and others. Though Molzym didn't disclose the cost of its instrument, the company anticipates that its kits would enable testing for bacterial and fungal targets at less than $100 per test.
"There’s no doubt that fast, accurate, and actionable information for patients with suspected acute infections and sepsis is a major unmet need … [and] Molzym is continuing to push the boundary on better diagnostics," said Tim Sweeney, cofounder and CEO of Burlingame, California-based startup Inflammatix, which is developing a host-response molecular test for rapid sepsis diagnosis. "Sequencing is clearly going to be a relevant addition to molecular detection approaches in certain patient subgroups with severe infections, especially the immunocompromised, where panel approaches can fail to detect rarer organisms," said Sweeney, who is not affiliated with Molzym.
"That said, any overnight test — or really any test that takes longer than about 30 minutes — is likely going to be useful mostly for antibiotic narrowing or, occasionally, broadening, since the decision to order antibiotics is usually made in the first hour," he added.
Lustig said he believes that by marketing a test that operates directly from patient samples and provides broad pathogen coverage, Molzym will be able to differentiate itself from other companies in the market for sepsis testing. For current sales in Europe, the firm uses a network of distributors, but for future US sales post-clearance, the firm is planning to use an internal sales team and work in parallel with distributors to target hospitals and private reference laboratories.
Established diagnostic companies ─ including Roche, BioMérieux, Accelerate Diagnostics, T2 Biosystems, and others ─ are marketing products that aim to enable rapid diagnosis of sepsis. Except for T2 Biosystems, which markets direct-from-blood molecular diagnostic panels, all of these companies have testing products that detect sepsis after the sample has been cultured.
However, companies continue to develop more advanced products. For example, Inflammatix is developing a 30-minute gene expression-based immune response test, InSep, that distinguishes viral from bacterial infections and is targeted toward clinicians diagnosing sepsis. The firm plans to launch InSep, formerly called HostDx Sepsis, as a cartridge-based rapid isothermal assay running on its Myrna platform during the second half of this year.
Accelerate announced recently that it intends to introduce a next-generation platform, Pheno 2.0, that would enable it to enter the high-volume microbiology segment of the identification and antimicrobial susceptibility testing market by working with many sample types including blood, urine, and isolates. Its current system operates from blood after a positive culture result.
"It seems likely that future workflows will start with biomarker testing to determine which patients are likely to have bacterial infections and then move only those patients into direct-from-sample testing," Sweeney said. "In other words, start up front by answering whether the patient needs antibiotics, and then try to answer which antibiotics with ID and AST testing. This can keep costs in check while also getting critical, actionable information to physicians at the right time."