Huazhong University of Science and Technology Union Hospital Licenses ARMS-PCR-Based BCR-ABL Kinase Domain Mutation Detection Kit for CML to Xiamen Zhishan Biotech for RMB 200,000

Recently, Union Hospital, Tongji Medical College of Huazhong University of Science and Technology, released a public notice on the transformation of scientific and technological achievements, stating that the hospital intends to“Nucleic Acid Composition and Kit for Detecting ABL Kinase Domain of BCR-ABL Fusion Gene by ARMS-PCR”The invention patent is granted to Xiamen Zeesan Biotech Co., Ltd. under a non-exclusive license, with the licensing fee amounting to RMB200,000 yuan。

The inventor of this patented technology is the Department of Hematology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology.Professor Deng Jun's Team。

Prof. Deng has long been dedicated to research in molecular diagnosis and targeted therapy for hematologic disorders, accumulating extensive expertise in the detection of leukemia fusion genes, mutation screening, and clinical translation.

The assignee of this invention patent isXiamen Zeesan Biotech Co.,Ltd.Xiamen Zeesan Biotech Co., Ltd. is a high-tech enterprise integrating the R&D, production, sales, and service of molecular diagnostic reagents and instruments. Zeesan Biotech is committed to researching globally leading molecular diagnostic technologies and providing innovative total solutions to facilitate early disease detection, precision treatment, and prognosis monitoring.

The technical invention proposed for transfer in this instance puts forwardA nucleic acid composition and kit for detecting the ABL kinase domain of the BCR-ABL fusion gene by ARMS-PCR;The nucleic acid composition comprises primer and probe sequences designed based on the 19 common mutations in the ABL kinase domain of the BCR-ABL fusion gene and the ABL internal reference. The kit enables detection to be completed in 11 wells, featuring simple operation and a detection sensitivity of up to 1%. Compared with other common molecular biology detection methods, it significantly shortens the testing cycle and reduces experimental costs, while improving both sensitivity and accuracy.

Chronic Myeloid Leukemia (CML) is a malignant tumor of the hematopoietic system. The vast majority of CML patients (more than 95%) carry a so-calledBCR-ABLofFusion GeneThis gene is formed by the translocation and fusion of the ABL gene on human chromosome 9 with the BCR gene on chromosome 22.

The protein encoded by this fusion gene exhibits abnormally highTyrosine Kinase Activity, this activity continuously emits signals that promote cell growth and division, directly driving the transformation of normal cells into leukemia cells, thus playing a central role in the pathogenesis of CML.

The BCR-ABL fusion gene is not a single entity; variations in the specific breakpoints and fusion sites result in fusion proteins of different sizes, which are associated with distinct disease phenotypes.

There are three common types:The first type is the M-bcr type., with the breakpoint located in a specific region of the BCR gene, resulting in a fusion protein with a molecular weight of approximately 210 kDa. This subtype accounts for the vast majority of CML patients and approximately one-third of patients with Philadelphia chromosome-positive acute B-lymphoblastic leukemia (Ph+ B-ALL).The second type is the m-bcr type., resulting in a ~190 kDa fusion protein, which is primarily observed in the remaining two-thirds of Ph+ B-ALL patients and a very small subset of CML patients.The third type is the μ-bcr type,Produces a fusion protein of approximately 230 kDa, which is primarily associated with chronic neutrophilic leukemia.

The emergence of drugs targeting the tyrosine kinase activity of the BCR-ABL protein, namely tyrosine kinase inhibitors (TKIs, such as imatinib), has revolutionized the treatment landscape for chronic myeloid leukemia (CML). However, clinical resistance to TKI therapy rapidly emerged. Studies have identified that one of the key mechanisms underlying this resistance isPoint mutations have occurred in the ABL kinase domain of the BCR-ABL fusion gene.

These mutations alter the spatial conformation of the kinase domain, hindering the binding of TKI drugs and thereby diminishing or completely abolishing their efficacy. To date, more than 90 resistance-associated mutation sites have been identified, among which certain mutations (such as T315I) confer resistance to nearly all existing TKI drugs, resulting in a poor prognosis for patients.

In the same patient, the mutation types in the ABL kinase domain may undergo dynamic evolution at different stages of disease progression. Initially, mutant clones may be present at a low proportion, but once they emerge, they can expand rapidly, leading to disease progression. Even during treatment, original mutant clones may disappear, while new resistant clones may emerge.

Drug-resistant mutations frequently emerge during the blast phase of the disease; therefore, regular and highly sensitive monitoring of ABL kinase domain mutation status is crucial. This enables clinicians to promptly detect signs of drug resistance, providing a critical basis for adjusting the type or dosage of tyrosine kinase inhibitor (TKI) therapy and implementing personalized treatment. For instance, early detection of certain specific mutations when the mutational burden is still low holds greater positive clinical significance for guiding management.

Currently, the mainstream methods for clinical detection of ABL kinase domain mutations includeNested PCR MethodandNext-Generation Sequencing (NGS). Nested PCR enhances specificity through two rounds of amplification; however, the procedure requires opening the tubes containing the first-round amplicons to initiate the second round, which readily generates aerosol contamination and leads to false-positive results.

Meanwhile, the first-generation sequencing methods typically used in follow-up testing have limited sensitivity and struggle to detect low-frequency mutations. Although next-generation sequencing (NGS) has become increasingly prevalent in recent years and can significantly enhance detection sensitivity, its application is somewhat constrained in scenarios requiring rapid reporting to guide clinical medication due to higher experimental costs, operational complexity, and longer turnaround times.

In direct response to the practical pain points in current clinical testing, including insufficient sensitivity, susceptibility to contamination during operation, lengthy turnaround times, and high costs, this patented invention has developedARMS-PCR Detection Technologyemerged as the times required.

The ARMS-PCR detection technology proposed by this invention demonstrates its core advantages and advanced nature primarily in itsHigh-Specificity Detection PrincipleAbove. Based on the principle of Amplification Refractory Mutation System (ARMS), this technology employs carefully designed specific primers and probes targeting 19 high-frequency mutation sites within the ABL kinase domain of the BCR-ABL fusion gene.

The 3’ terminal bases of these primers perfectly match the target mutant sequence, thereby enabling efficient initiation and extension during amplification. In contrast, for the wild-type sequence, the mismatch at the terminal base significantly reduces the extension efficiency of DNA polymerase, thus achieving selective amplification of mutant versus wild-type sequences. This design allows the technique to specifically capture and amplify low-abundance mutant sequences against a background of abundant normal genes, laying the methodological foundation for high-sensitivity detection.

In specific detection systems, this technology demonstratesHigh-Efficiency Integrationcharacteristics. Through rational design of primer and probe combinations, the detection of 19 mutations is ingeniously integrated into 11 independent reaction tubes. Among these, some reaction tubes can simultaneously detect 2 to 4 different mutations. This multiplex detection design significantly enhances the throughput of a single test, reducing reagent consumption and sample volume.

CooperationReal-time Quantitative PCR Platform, the entire detection process is conducted under closed-tube conditions, thoroughly eliminating the risk of aerosol contamination associated with traditional nested PCR due to tube opening for the second round of amplification, thereby significantly enhancing laboratory operational safety and result reliability.

Another major advancement of this technology lies in itsExcellent Detection Performance and Convenience. The optimized reaction system and amplification protocol enable the entire process, from nucleic acid extraction to result output, to be completed within 3 to 5 hours, with a detection sensitivity of up to 1%. This means that even if mutant cells constitute only 1% of the cell population, they can be effectively detected. This is of critical significance for the early clinical detection of low-level resistant clones and timely therapeutic intervention.

Meanwhile, the entire testing procedure is standardized, with primary instrument requirements limited to conventional real-time quantitative PCR instruments. This facilitates its implementation in clinical molecular diagnostic laboratories of most hospitals, thereby greatly enhancing the accessibility of this technology.

Compared with existing mainstream technologies, this patented technology has achieved a favorable balance and significant breakthroughs. In contrast to nested PCR combined with Sanger sequencing—a method characterized by cumbersome operations, susceptibility to contamination, and limited sensitivity—this technology is faster, safer, and more sensitive. Compared with next-generation sequencing (NGS), which is regarded as the gold standard, this technology substantially reduces testing costs and shortens the turnaround time while ensuring the clinical-level sensitivity required for detecting target high-frequency mutations, thereby better meeting the urgent clinical need for rapid reporting to guide medication.

Therefore, this patented technology provides an accurate, rapid, cost-effective, and easy-to-implement screening protocol for ABL kinase domain mutations, demonstrating significant potential for clinical translation and application.

In response to the urgent demand in today’s precision medicine market for multi-target, integrated testing solutions, relevant enterprises and medical institutions are advancing the research and development of various molecular diagnostic kits for other hematologic diseases and solid tumors, building upon their deep expertise in leukemia diagnosis and treatment pipelines. Their goal is to establish a comprehensive product ecosystem covering the entire process of screening, diagnosis, and prognosis monitoring.

In the international market,Terns PharmaceuticalsIn the field of targeted therapy for chronic myeloid leukemia (CML), the company has developedTERN-701, a Next-Generation Allosteric BCR-ABL Inhibitor. This drug is an oral, potent, and highly selective allosteric tyrosine kinase inhibitor. Its mechanism of action involves targeting the myristoyl pocket of the BCR-ABL protein, thereby enabling effective treatment of chronic myeloid leukemia (CML), particularly in patients who are resistant to or have suboptimal responses to existing therapies, including other allosteric inhibitors.

Currently, TERN-701 is in an accelerated phase of clinical development. Its CARDINAL Phase I/II clinical trial for chronic myeloid leukemia (CML) is ongoing and progressing smoothly. The drug received Fast Track designation from the U.S. FDA in the fourth quarter of 2025.

In China,Haixi Biotechnology Co., Ltd.Jointly established by the Medical School of Wuhan University and KingMed Diagnostics Gene Technology Co., Ltd. The company primarily focuses on leveraging advanced molecular diagnostic technologies, such as high-throughput sequencing, to provide integrated solutions for the differential diagnosis, classification, prognostic assessment, and targeted therapy guidance of hematologic malignancies. In the field of targeted therapy support for chronic myeloid leukemia (CML), Haixi Biotech has successfully developed and launched“Human 60-Gene Fusion Screening Kit”。

This kit can simultaneously detect 60 fusion genes associated with 241 leukemia subtypes, including the BCR-ABL fusion gene and its variants, which are critical for the diagnosis and treatment monitoring of chronic myeloid leukemia (CML). The product is now commercially available and has been applied in clinical diagnostics.

Looking Ahead at the Industry’s Future, Both Diagnostics and Therapeutics Continue to Evolve.

At the therapeutic level, targetingTargeted therapy represented by tyrosine kinase inhibitors (TKIs) of BCR-ABLIt remains the cornerstone of chronic myeloid leukemia (CML) treatment, while next-generation drugs targeting refractory mutations such as T315I and novel therapies like PROTACs, which directly degrade pathogenic proteins, are under development to further overcome drug resistance. In terms of detection, the advancement of precision medicine has imposed higher demands on the sensitivity, speed, and cost-effectiveness of mutation testing.

In the future, more sensitive digital PCR (dPCR) and next-generation sequencing (NGS) technologies capable of scanning a broader range of mutation sites will complement existing methods, jointly serving clinical practice.