Jilin University Licenses Novel Trichomonas tenax Detection Technology Based on GAPDH Antigen to Changchun Booude Biotech for RMB 1.32 Million

Recently, Jilin University released a public notice on the transformation of scientific and technological achievements, proposing to transfer relevant technologies through agreed pricing.“Trichomonas vaginalis Antigenic Gene Protein Glyceraldehyde-3-Phosphate Dehydrogenase and Medical Uses Thereof”Relevant patents have been assigned to Changchun Bio-Europe Biotechnology Co., Ltd., with the assignment amount beingRMB 1.32 million. The inventors of this patent areProfessor Zhang Nan and his team。

This technology, developed by Jilin University, is based onImmunological Detection Technology Using Trichomonas vaginalis Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH) as a Specific Antigen, which is primarily used for the precise detection of *Trichomonas vaginalis* infections. It has also completed the screening and preparation of the antigen protein, as well as established the foundational framework for the development of related diagnostic kits. This represents a technological achievement at the intersection of genetic engineering and immunology.

As a zoonotic pathogenic protozoan, *Pentatrichomonas hominis* infection is closely associated with diarrhea and intestinal barrier damage, and is highly correlated with the pathogenesis of colorectal cancer. With a broad host range encompassing humans, companion animals, and livestock, the demand for rapid, accurate, and convenient detection methods for *P. hominis* continues to rise in both clinical and veterinary fields as understanding of its pathogenicity deepens.

However, existing detection technologies suffer from numerous core pain points that severely compromise the efficiency of infection screening and the accuracy of diagnosis, thereby failing to meet the diverse testing demands under the prevention and control of zoonotic diseases.

Traditional detection technologies face core diagnostic bottlenecks; on one hand,Direct smear microscopy has a low detection rate and is prone to missed or misdiagnosis,Although this method is simple to operate and does not require specialized instruments, it relies on manual microscopic observation of the parasites. It is highly influenced by the examiner’s experience, parasite load, and sample processing methods. It is prone to false negatives in samples with low parasite burdens, cannot be effectively differentiated from other Trichomonas species, and has a high misdiagnosis rate, making precise diagnosis difficult. On the other hand,High Operational Threshold of PCR Testing Limits Its Application in Various ScenariosAlthough this method offers high sensitivity and strong specificity, it requires specialized equipment such as PCR instruments and real-time fluorescence quantitative detectors. Furthermore, it imposes stringent requirements on the experimental environment and the professional competence of operators, restricting its use to specialized laboratories. Consequently, it cannot achieve rapid detection in settings such as primary healthcare institutions or livestock farming sites, thereby hindering the implementation of on-site screening and point-of-care diagnosis.

From the perspective of practical application scenarios, existing technologies still exhibit significant limitations in utility. Traditional detection methods lack standardized immunological testing protocols; the interpretation of smear microscopy results has no unified quantitative standards and relies entirely on subjective manual judgment, leading to poor inter-operator consistency. The PCR detection method not only involves cumbersome sample pre-processing and long turnaround times but also suffers from high reagent costs and a propensity for false-positive results. Furthermore, it cannot facilitate rapid screening of batch samples, resulting in extremely low efficiency in large-scale epidemiological investigations and livestock population testing.

Meanwhile, existing diagnostic workflows are complex. Smear microscopy involves multiple steps, including sample collection, slide preparation, staining, and microscopic examination, while PCR testing requires nucleic acid extraction, amplification, and result analysis. These processes are time-consuming and fail to meet the demand for point-of-care testing in scenarios such as clinical emergencies and rapid on-site screening in aquaculture, thereby delaying infection control and treatment.

Furthermore, existing technologies have failed to fill the market gap in immunological testing. Currently, there are no specific detection antigens or standardized immunological assays for *Pentatrichomonas hominis*, nor are there any rapid-test kit products available. Traditional detection methods fail to balance sensitivity with convenience and struggle to meet the needs of both professional laboratory testing and point-of-care testing at primary care levels. While direct smear microscopy is convenient but lacks precision, and PCR testing is accurate but limited in application scenarios, neither method enables non-invasive serological testing, creating a blind spot for patients from whom fecal samples cannot be collected.

Meanwhile, the absence of unified quality control standards for existing detection technologies, coupled with variations in testing methods and interpretation criteria across different laboratories, results in a lack of comparability among test outcomes. This hinders cross-regional prevention and control efforts as well as epidemiological research on *Tritrichomonas hominis* infection. These issues collectively have led to challenges in the detection of *Tritrichomonas hominis*.“Microscopy is prone to missed diagnoses, PCR has a high threshold, and there is no convenient immunoassay solution”dilemma, there is an urgent need in both clinical and veterinary fields for a highly sensitive and specific, easy-to-operate, and scenario-adaptive testing solution to address pain points across the entire workflow from sample detection to practical application.

Addressing the three core pain points in the detection of *Trichomonas hominis*—namely, the high rate of missed diagnoses with microscopy, the high technical barrier for PCR operations, and the lack of standardized immunological testing protocols—the research and development team at Jilin University has innovatively launchedImmunological Detection Technology Using Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH) as a Specific Antigen, with"Highly Sensitive and Specific Core Antigens + Convenient and Broad-Spectrum ELISA Testing"As its core advantage, it establishes a full-process solution ranging from antigen preparation to detection applications, breaking through the clinical and scenario limitations of traditional detection technologies, and providing a new technological paradigm for the precise screening and efficient diagnosis of Trichomonas hominis infection.

This technology pioneers the screening of highly immunogenic, specific detection antigens., filling the gap in industry-specific antigens. The R&D team precisely screened and identified GAPDH protein as the core detection antigen from the excretory/secretory antigens of *Pentatrichomonas hominis* using LC-MS/MS mass spectrometry analysis. Its gene sequence is clearly defined (SEQ ID NO: 1), and experimental verification has confirmed its localization in the cytoplasm, on the surface, and within the flagella of the parasite, demonstrating excellent immunogenicity. A recombinant plasmid was constructed using the pET-32a expression vector, enabling high-level expression in *E. coli* BL21 cells. High-purity GAPDH protein was obtained through nickel column purification, laying a highly specific material foundation for immunological detection and fundamentally changing the status quo of lacking exclusive detection antigens for *Pentatrichomonas hominis*.

Meanwhile,This technology establishes an optimized indirect ELISA detection system, enabling highly sensitive and specific standardized testing.Through checkerboard titration, comprehensive optimization of all dimensional conditions was performed to determine core parameters such as optimal antigen coating concentration and serum dilution. The developed ELISA assay achieved a sensitivity of 1:160, enabling accurate detection even when positive sera were diluted 160-fold. It exhibited excellent specificity with no cross-reactivity against related parasites such as Tritrichomonas foetus, Toxocara canis, and Giardia lamblia. The intra-assay and inter-assay coefficients of variation were less than 7% and 8%, respectively, demonstrating superior reproducibility. Furthermore, clearly defined cutoff values for positive and negative results enabled standardized, quantitative interpretation, thereby thoroughly addressing the issues of subjective judgment, missed diagnoses, and misdiagnoses associated with traditional smear microscopy.

Furthermore,This technology has established a convenient, broad-spectrum testing modality, overcoming the limitations of scenario-specific applications.Compared with the stringent requirements of PCR testing for specialized instruments and laboratory environments, this ELISA-based assay is simple to operate, requiring only conventional reagents and basic equipment. It can be performed after minimal training, features a short turnaround time, and enables rapid high-throughput screening. The use of non-invasive serum sampling enhances patient acceptance and is suitable for individuals from whom fecal samples are difficult to collect, thereby significantly broadening its applicability. Furthermore, it demonstrates 100% concordance with traditional fecal examinations, fully validating its clinical accuracy and reliability. This method meets the diagnostic needs across diverse settings, including primary healthcare institutions, veterinary stations, pet hospitals, and large-scale epidemiological surveys.

At the level of clinical promotion and industrial value,This technology offers the advantages of standardized mass production and low-barrier widespread adoption.The core GAPDH antigen can be standardized for mass production, enabling the preparation of diagnostic kits for *Pentatrichomonas hominis* and facilitating industrial-scale manufacturing and widespread adoption. The detection method does not require high-end specialized equipment, significantly lowering the barrier to testing and making it suitable for medical and livestock testing institutions at all levels, thereby filling a market gap for immunological diagnostic kits for *P. hominis*. From a public health perspective, this technology provides a practical technical solution for the early screening, diagnosis, and prevention and control of *P. hominis*, a zoonotic protozoan parasite. It can effectively interrupt the fecal-oral transmission route, reduce the risk of cross-species infection between humans and animals, and provide data support for colorectal cancer-related risk screening and infection source tracing, thereby promoting the development of *P. hominis* detection toward greater precision, efficiency, and accessibility.

Currently, traditional technologies remain the mainstream in the market for Trichomonas vaginalis detection, while universities and research institutions are focusing on the development of immunological detection technologies, forming“Scaled Application of Traditional Methods + Breakthroughs in Innovative Immunoassay Technologies”...competitive landscape. Key participants include biotechnology companies specializing in parasitic detection, university research teams, and clinical laboratory institutions. Technological approaches focus on optimizing traditional microscopy, improving PCR techniques, and screening for specific antigens, leading to differentiated development in terms of detection accuracy, operational convenience, and scenario adaptability. Among these, innovative immunological assays are mostly in the stage of laboratory validation or technology transfer.

Jiangxi Agricultural University Research TeamInventedA Method for Detecting Trichomonas hominis Based on the RPA-CRISPR-Cas12a SystemThis method relies on crRNA designed for specific gene fragments of *Tritrichomonas hominis* and upstream and downstream primers used for recombinase polymerase amplification (RPA) of nucleic acids. It achieves, for the first time, the differentiation between *Trichomonas fetus* and *Tritrichomonas hominis* based on the RPA-CRISPR-Cas12a system, providing a robust technical approach for grassroots monitoring and detection of *Tritrichomonas hominis* and *Trichomonas fetus* infections. The detection method based on the RPA-CRISPR-Cas12a system is performed under isothermal conditions, requires no specialized instrumentation, and enables rapid and accurate detection in field settings, demonstrating high sensitivity and specificity.

Team of Li Wenchao, Anhui Science and Technology UniversityDevelopedA Duplex Nested PCR Method for Simultaneous Detection of Tritrichomonas foetus and Tritrichomonas blagburni, and a patent for invention has been filed for the related technology. The team designed specific primers based on the SSU rRNA gene of *Pentatrichomonas hominis*. Through a two-round nested PCR amplification process, the first round uses genus-specific primers for *Trichomonas*, and the second round employs a mixture of species-specific primers. This method can simultaneously detect *Pentatrichomonas hominis* (target fragment: 339 bp) and *Tetratrichomonas buttreyi* (target fragment: 623 bp). Experimental validation demonstrated that the detection sensitivity reaches 500 fg/μL, with no cross-reactivity against various other protozoa such as *Tritrichomonas foetus* and *Giardia*. Furthermore, the results show good concordance with those from single-target nested PCR assays, significantly reducing detection time and costs. Currently, this technology has completed clinical validation using 300 bovine fecal samples, making it suitable for simultaneous screening and epidemiological surveys of these two trichomonad species in cattle farming settings.

The immunological detection technology for the *Trichomonas hominis* GAPDH protein antigen, developed by Jilin University, establishes a highly sensitive and specific proprietary testing solution that is easy to operate. Compared with traditional microscopy, universal nucleic acid testing, and the duplex nested PCR technology developed by Anhui Science and Technology University—which can simultaneously detect two species of *Trichomonas*—this method demonstrates significant advantages in specificity, convenience, and adaptability to various scenarios. It provides core technical support for the screening and detection of this parasite as well as for the development of related test kits, holding substantial clinical and public health value.