Objective: To systematically elaborate the design and clinical trial requirements of respiratory multiple pathogen nucleic acid detection kits in China, so as to provide a reference for the development and clinical trials of related products. Methods: The approval overview of respiratory multiple pathogen were summarized. Combining the expected clinical situation and related diagnostic consensus, an in-depth analysis was conducted on the considerations in technical review process of relevant products from the aspects of the combination of pathogens covered, the applicable sample types, the expected applicable population, and clinical trial requirements. Results and Conclusion: Respiratory multiple pathogen nucleic acid detection kits play a significant role in the auxiliary diagnosis of respiratory tract infections. The product design should align with intended use, and the principal investigators of clinical trials should combine the design and intended use of the products, thoroughly studying the enrolled population, comparative methods, number of cases, and statistical analysis to evaluate the clinical performance of the products more scientifically and reasonably, and facilitate timely regulatory approval.

Objective: To provide reference for the research and development direction and product marketing of in vitro diagnostic reagents for dengue virus detection. Methods: The current clinical diagnostic methods for dengue virus infection in China were elaborated, the approved in vitro diagnostic reagent products were sorted out, and the research and development direction and pre-marketing regulatory requirements of dengue virus detection in vitro diagnostic reagents were analyzed based on the clinical diagnostic needs of the disease and the market situation of related products. Results and Conclusion: There are multiple methods for laboratory testing of dengue virus, each with its own advantages and limitations. At present, the National Medical Products Administration has approved a total of 11 products, of which 9 are antigen, antigen antibody combination, antibody immunological testing methods, and the other 2 are nucleic acid testing. However, there are few highly sensitive and specific nucleic acid testing kits available, which cannot meet clinical needs. It is recommended that relevant production enterprises increase their efforts in developing such products and conduct product evaluations in a scientific and reasonable manner. Product development should be based on the clinical diagnostic needs of the disease, and product evaluation should be scientifically and reasonably evaluated based on specific conditions such as the expected use of the product, the detected biomarkers, and the expected target audience. The clinical performance of the product should be fully evaluated to promote its rapid marketing.

Objective: To study the establishment and optimization of the pathogen targeted next-generation sequencing (tNGS) process based on the probe capture method, as well as the automated implementation of it, and to verify the analytical performance. Methods: The factors influencing the tNGS methodology were analyzed, including the probe design and production quality control, data volume and sequencing read length. The analytical performance was evaluated by using a reference panel constructed with 62 types of microbes, including various bacteria, fungi, viruses, and atypical pathogens. As well as, the differences in limit of detection, precision and cross-contamination between manual operations and automated were compared. Results: The study showed that as the probe coverage increased, the limit of detection performance increases accordingly, and the probe still maintained high specificity. The optimized tNGS demonstrated good analyze performance in terms of accuracy, limit of detection, precision, specificity linearity and so on. Under the premise of the same limit of detection performance, the automated method was superior to manual operation in precision and cross-contamination resistance performance. Conclusion: The tNGS process established in this study has good analytical performance and provides a new strategy for clinical pathogen detection. Future studies should focus on clinical sample confirmation and automated process validation to promote better application of the technology in practical clinical work.

Objective: To carry out the research on the consistency evaluation of human papillomavirus(HPV) nucleic acid detection ability, evaluate the HPV nucleic acid detection proficiency of different domestic laboratories, so as to promote the mutual recognition of HPV nucleic acid detection results in domestic laboratories. Methods: HPV samples containing 14 different high-risk types (HPV-16, -18, -31, -33, -35, -39, -45, -51, -52, -56, -58, -59, -66 and -68) and 2 low-risk types (HPV-6, -11) were prepared with both type specificity and quantitative accuracy. Then panel was sent to different laboratories in the form of blind samples for testing. The proficiency study results were evaluated by the method of total integral grading evaluation. Results: A total of 27 laboratories participated in this proficiency study, of which 22 laboratories’ test results were “satisfactory” and 5 laboratories’ test results were “unsatisfactory”, with a satisfaction rate of 81.5%. Conclusion: The capacity of HPV test in different laboratories varies greatly, and the detection ability of some laboratories is poor. It is recommended to analyze the causes, strengthen laboratory training and timely rectification from four aspects: personnel, equipment, kit and quality control, so as to improve their laboratory testing capabilities.

Objective: To evaluate 5 kinds of domestic anti-HCV Colloidal gold detection kits on the market, so as to provide reference for selecting antibody screening test kits. Methods: 200 negative samples and 141 positive nucleic acid samples were tested with 5 kinds of domestic rapid diagnostic reagents for HCV antibody. Results: The false positive and false negative rates of GZWF were both 0%; the false positive rate of YKXC was 1%, and the false negative rate was 1.4%; the false positive rate of BJWT was 2%, and the false negative rate was 1.4%; the false positive rate of SHKH was 0%, and the false negative rate was 0.7%; the false positive rate of HZAB was 1%, and the false negative rate was 0%. Conclusion: The sensitivity of five domestically produced HCV antibody colloidal gold test kits ranges from 98% to 100%, and the specificity is 98.5%-100%. The sensitivity and specificity are greater than 98%. The overall quality is relatively high, and the results are reliable, which can serve as a rapid and efficient screening method for detecting hepatitis C infected individuals.