Key points of novel coronavirus detection reagent registration technology review points issued by the drug administration

Novel coronavirus antigen / antibody test registration technical review points (Trial) were released on 25 th by the state medical and pharmaceutical regulatory commission's medical device technology evaluation center 2019 Novel coronavirus antigen / antibody detection reagent registration information was prepared and written by registered applicants, and it also provided reference for the technical review department to review registration data 2019 The novel coronavirus antigen / antibody detection reagent is generally required for 2019 new types of coronavirus Applicants should determine whether the content is applicable according to the specific characteristics of the product If not applicable, the reasons and corresponding scientific basis should be specified and the contents of the registration declaration materials should be substantiated and refined according to the specific characteristics of the products The novel coronavirus novel coronavirus antigen / antibody detection reagent is applied to 2019 new coronavirus antigen / antibody detection reagent The reagent is used for the qualitative detection of 2019 new coronavirus antigen / antibody in serum, plasma, whole blood, throat swab, bronchoalveolar lavage fluid, sputum or other respiratory secretions The review points are applicable to the products for which the first registration application is made The performance evaluation enterprise shall submit all the research data of performance evaluation and verification of the kits produced under the production environment conforming to the quality management system, including the specific research method, experimental scheme, experimental data, statistical analysis and other detailed data It is suggested to focus on the following analysis performance 1 Sample collection and processing (1) the selection of time point for sample collection: whether it is affected by the course of disease, clinical symptoms, medication and other factors (2) For swab samples, the requirements for sampling swab and sample preservation solution (sampling solution) are studied: the requirements for the material of sampling swab (including the requirements for swab head and swab rod), the requirements for preservation container, the requirements for the composition, concentration and usage of preservation solution (sampling solution, etc.) (3) For blood samples, the suitable anticoagulants were studied 2 Minimum detection limit (1) determination of minimum detection limit Antibody detection: it is recommended to select specific antibody positive samples of specific titer for gradient dilution to determine the minimum detection limit Each gradient of diluent is repeated for 3-5 times, and each diluent is repeated for at least 20 times The antibody level with 90% - 95% positive detection rate is taken as the lowest detection limit, and the antibody titer should be confirmed by appropriate methods Antigen detection: it is recommended to study and determine the minimum detection limit after gradient dilution of virus, repeat 3-5 copies of each gradient of virus diluent, and repeat detection for at least 20 times for each diluent, take the virus level with 90% - 95% positive detection rate as the minimum detection limit, and confirm the antigen concentration with appropriate methods Three representative clinical samples from different sources or virus stock solution after culture should be selected to determine the minimum detection limit of antigens, IgM antibodies and IgG antibodies respectively (2) At least 3 virus strains or clinical samples (different from the samples determined by the minimum detection limit) with time and regional characteristics shall be selected for the verification of the minimum detection limit at the concentration level of the minimum detection limit, and the positive detection rate shall reach 90% - 95% The diluent used shall be consistent with the matrix of the applicable sample type and may be diluted with a negative sample The antigen detection reagent shall provide a detailed method to determine the virus titer, and at the same time, it shall describe the confirmation method and verification results of virus samples in detail Antibody detection reagent shall provide detailed confirmation method and results of antibody type and titer 3 The verification of the tolerance of virus samples in different regions provides real clinical samples of patients from different sources with time and regional characteristics for verification Antigen, IgM antibody and IgG antibody detection reagents are used to study 10 different patient samples respectively The verification content should include the minimum detection limit, repeatability, etc., and provide the confirmation method and experimental data of samples and concentrations 4 Analysis specificity (1) cross reaction verification (antigen, IgM antibody and IgG antibody detection reagent should be verified respectively) ① local human coronavirus (hku1, OC43, nl63 and 229E); ② H1N1 (new H1N1 influenza virus (2009), seasonal H1N1 influenza virus), H3N2, H5N1, h7n9, influenza B Yamagata, Victoria, respiratory syncytial virus, rhinovirus a, B, C, adenovirus 1, 2, 3, 4, 5, 7, 55, enterovirus a, B, C, D, EB virus, measles virus, human cytomegalovirus, rotavirus, norovirus, mumps virus Varicella zoster virus; ③ Mycoplasma pneumoniae Provide the source, yin and Yang, species / type, concentration / titer confirmation and other test data of all pathogens, antibodies and other samples used for cross reaction verification Antigen detection reagents are recommended to conduct cross reaction verification at medical-related levels such as viruses, such as virus concentration of 105 PFU / ml or higher The novel coronavirus specific IgG antibody with high concentration was verified by cross reaction with specific IgM antibody ⑤ Verify no less than 20 normal human samples (2) The potential interferences of different types of samples may be different, so the suitable interferences should be selected according to the specific types of samples It is suggested that the applicant should evaluate the potential maximum concentration ("worst condition") of each interfering substance and carry out interference test verification at the critical positive level of virus antigen and antibody 5 IgM antibody detection reagent: carry out IgM destruction experiment on at least 5 samples containing pathogen specific IgM antibody The method is to treat the samples with specific chemicals (such as 2-mercaptoethanol or dithiothreitol) and carry out the detection again The IgM detection result should be negative 6 Precision enterprises shall make reasonable requirements for the evaluation standards of precision indexes, such as standard deviation or coefficient of variation The simulation sample can not reflect all the variation factors that the clinical sample may bring, so the precision evaluation should include several clinical samples at the same time The precision evaluation of this kind of products mainly includes the following requirements (1) Verify the main variables that may affect the detection precision, and verify the detection time, analyzer, operator, location, test rounds and other factors (2) Set a reasonable precision evaluation cycle, for example: testing for at least 20 days or 5 days, the specific scheme can refer to EP documents So the precision of in batch / inter batch, in day / day and different operators can be evaluated comprehensively (3) Clinical samples used for precision evaluation should contain at least three levels: negative samples, critical positive samples, and (medium or strong) positive samples, and appropriate precision requirements should be set according to product characteristics ① Negative sample: the concentration of the substance to be tested is lower than the minimum detection limit or zero concentration, and the negative detection rate shall be 100% (n ≥ 20) ② Critical positive sample: the concentration of the substance to be tested is slightly higher than the minimum detection limit of the kit, and the positive detection rate should be ≥ 95% (n ≥ 20) ③ Medium / strong positive sample: the concentration of the substance to be tested is medium to strong positive, the positive detection rate is 100% and CV ≤ 15% (n ≥ 20) 7 Hook effect 8 Provide enterprise reference product verification data: according to the enterprise reference product settings in the main raw material research data, use three batches of products to inspect the enterprise reference products and provide detailed experimental data 9 For other problems needing attention, such as the products using the instrument to interpret the results, the performance evaluation data of all models of instruments listed in the product manual [applicable models] shall be provided 10 Applicable sample type if the product is suitable for serum and plasma, homology comparison can be used to verify the comparability of the sample If the product is suitable for whole blood, the sample type can be used for full performance evaluation, at least for the minimum detection limit, the tolerance and precision of virus samples in different regions, and for homologous comparison test The registration inspection shall provide the product inspection report that meets the technical requirements of the product and is conducted in the medical device inspection institution with the corresponding medical device inspection qualification and acceptance scope, and shall provide the inspection qualification report of three consecutive production batches of samples There is no national reference for the project, so the enterprise reference should be used for registration inspection If the national reference for emergency use is issued, the national reference should be used for registration inspection The positive judgment value determination data shall be submitted to the research data determined by the positive judgment value (cut off, CO) of the declared reagent negative / gray area / positive results, including the specific test scheme, sample selection of population and subjects, evaluation criteria, statistical analysis and research data, etc The selection of sample sources used to establish positive judgment value should take into account the influence of different geographical areas, different infection stages and physiological conditions In addition, determine whether there are differences among different sample types If there are differences, determine them separately If applicable, the receiver operating characteristic curve (ROC) analysis method can be used to select and determine the reasonable positive judgment value; if the result has the gray area, the basis for establishing the gray area shall be clear If other methods are used to confirm the positive judgment value, the rationality of this method should be explained Main raw materials research data the main raw materials of this product include antigens, antibodies, quality control products, reference products, etc The selection and source of main raw materials, preparation process, quality standard and other relevant research data shall be provided If the main raw materials are self-made by the enterprise, the detailed preparation process shall be provided; if the main raw materials are sourced from outsourcing, the data shall be provided, including: the basis for selecting the raw materials, the comparative screening test data, the quality standards provided by the supplier, the delivery inspection report, and the quality inspection data after the arrival of the raw materials The supplier shall be fixed and shall not be replaced at will The applicant shall specify the quality control standard for each main raw material 1 novel coronavirus specific antigens, antibody specific pathogen antigens and antibodies are the key raw materials for such products Because the difference between novel coronavirus infected strains of different geographical and different populations is not clear, it is important to choose antigen and antibody raw material when selecting the antigen epitopes, avoid false negative differences among different strains, and also consider the expression of antigens in other coronary diseases, so as to avoid cross reaction The consideration of this aspect should be detailed in the research data of antigen and antibody raw materials (1) novel coronavirus antibody detection reagent specific antigen should first be detailed about antigen epitopes and selection basis In addition, the detailed test data of antigen source, preparation, screening, purification, identification and quality standard (appearance, protein concentration, purity, molecular weight, functional test, etc.) should be submitted It mainly includes the following two situations: ① if the self-made antigens are natural antigens, the experimental process of virus strain selection, culture, antigen extraction, purification and identification should be described in detail If it is a recombinant antigen, detailed information about specific gene selection, sequence information, Clone Construction and transformation, antigen expression, purification and identification shall be submitted ② The name of the antigen, the biological source of the antigen, the name of the supplier, the research data selected by the supplier and the performance index and test report of the antigen issued by the supplier shall be detailed (2) novel coronavirus antigen detection reagent specific antibodies should first be detailed about the epitopes targeted by antibodies, antibodies used to prepare the immunogen and determine the antibody.