40 results about "Firefly Luciferases" patented technology

The invention discloses a group of chicken PGC-1alpha gene promoters as well as a construction method and application thereof, and belongs to the field of gene engineering. The chicken PGC-1 alpha gene promoter is any one of A1-A5 sequences of which the nucleotide sequences are shown as SEQ ID NO: 1 to SEQ ID NO: 5. The promoter is applied to the promotion of expression of firefly luciferase in chicken preadipocytes, and is applied to the regulation and control of the chicken PGC-1 alpha gene promoter by a specific transcription factor PPAR gamma. The five PGC-1alpha gene promoters have promoter activity in chicken preadipocytes, and can directly regulate and control transcription expression of the chicken PGC-1alpha gene. The chicken PGC-1alpha promoter disclosed by the invention can predict the transcriptional activity of the PGC-1alpha at the in-vitro cellular level, so that the chicken PGC-1alpha promoter has potential application values for revealing a transcriptional regulation mechanism of the chicken PGC-1alpha, breeding low-abdominal-fat broilers, revealing molecular mechanisms after various human diseases occur and developing drugs for targeting PGC-1alpha expression.

The invention discloses a construction method and application of a rabies virus RNA polymerase dual-luciferase report system, and belongs to the technical field of biology. In order to solve the technical problem that a report system for detecting the RABV RNA polymerase activity needs to be developed urgently, a Firefly luciferase fragment and a vector pcDNA3.1-SRV9-M-G obtained by amplifying M protein and G protein-deleted rabies virus SRV9 strain full-length plasmid are seamlessly connected, a recombinant luciferase report plasmid is constructed, the recombinant luciferase report plasmid and an auxiliary plasmid are combined, and the recombinant luciferase RNA polymerase activity detection system is constructed. And the RABV RNA polymerase dual-luciferase report system is formed by the Renilla luciferase report plasmid and the T7 promoter plasmid, and the RABV RNA polymerase dual-luciferase report system comprises the Renilla luciferase report plasmid and the T7 promoter plasmid. The system can accurately and efficiently detect the activity of RABV polymerase, and by virtue of the function, effective antiviral targets and therapeutic drugs can be screened out in an assisted manner.

The invention discloses a promoter sequence of a large yellow croaker SOCS3 gene. The promoter sequence is as shown in SEQ ID NO. 1. The recombinant plasmid pGL3-SOCS3 is used for transfecting a carp epithelioma (EPC) cell, renilla luciferase is used as an internal reference, and the activity of firefly luciferase can be 4.2 times of that of a control group at most, which shows that the large yellow croaker SOCS3 promoter promotes the transcription expression of a downstream reporter gene and has the basic activity of the promoter. Recombinant plasmids pGL3-SOCS3 and pRL-TK are co-transfected into EPC cells, then poly (I: C) is added, and the result shows that the poly (I: C) can significantly enhance the activity of the large yellow croaker SOCS3 promoter, and the activity of the large yellow croaker SOCS3 promoter is 1.7 times higher than that of a control group, indicating that the large yellow croaker SOCS3 promoter can respond to exogenous immunostimulation. The invention provides an excellent experimental system for studying the transcriptional regulation mechanism of the large yellow croaker SOCS3 gene, can also be used for efficiently expressing exogenous genes or studying transgenic large yellow croaker, and has theoretical significance and practical value.

The invention provides application of microbial metabolites in tumor immune response and an intestinal microbial metabolite screening method. A screening system is constructed by combining firefly luciferase with PD-1 / SHP-2, various intestinal microbial metabolites are independently co-cultured with the screening system, the screened microbial metabolites are glycodeoxycholic acid and taurocholic acid, and the influence of various metabolites on PD-1 pathway signal transduction is reflected by measuring the activity of luciferase. The invention constructs a reliable, rapid and effective screening method for discovering intestinal microbial metabolites influencing a PD-1 signal channel and evaluating the influence efficacy of the metabolites, and screening out metabolites having effects on intestinal cancer immune response, thereby providing an experimental basis for enhancing intestinal cancer treatment.

This invention provides a method for constructing a visualized mouse model characterizing hepatocyte necrosis and apoptosis, belonging to the field of biomedical technology. The invention involves disassembling firefly luciferase (Fluc) into inactive N-terminal (Nluc) and C-terminal (Cluc) fragments, which are then fused and expressed at specific locations on the MLKL molecule, forming two independent plasmid vectors, MLKL-Nluc and MLKL-Cluc. These two plasmid vectors are then injected into mouse hepatocytes via tail vein high-pressure hydrodynamic injection to construct the visualized mouse model characterizing hepatocyte necrosis and apoptosis. This invention allows for non-invasive and dynamic monitoring of the execution process of necrosis and apoptosis—MLKL oligomerization—in animal models, providing a powerful and specific tool that fills the technological gap in real-time visual monitoring of necrosis and apoptosis in pan-apoptotic research.

The application belongs to the field of plant synthetic biology and plant gene expression regulation, and particularly relates to a vector for plant promoter double system quantitative characterization, a standardization method, a product and application. The method combines a protoplast transient expression system and an agrobacterium-mediated leaf transient expression system, uses a compact promoter characterization plasmid (pLIP) containing a firefly luciferase module driven by a to-be-detected promoter and a renilla luciferase internal reference module driven by a 35S promoter, and the two modules are located in a T-DNA interval. The ratio of the two luciferases is calculated, the RPU (relative promoter unit) method is introduced, a standard promoter (35S) is used as a reference, and normalization and direct comparison of promoter activities in different systems are realized. The scheme significantly improves the repeatability and cross-system compatibility of promoter activity data, and is suitable for efficient screening and standardized characterization of regulatory elements in plant synthetic biology.

The present invention belongs to the field of genetic engineering, and specifically discloses a novel firefly luciferase variant and its application in rapid detection of microbial content and as a reporter gene. The novel firefly luciferase mutant disclosed in the present invention has significantly improved catalytic activity and thermal stability, and the luminescent wavelength is easier to pass through the cell membrane and is more sensitive to low-concentration microorganisms. Therefore, the novel firefly luciferase variant is more suitable for rapid detection of microbial content and use as a reporter gene.

The application discloses a firefly-NanoLuc-AkaLuc three-luciferase reporter gene detection kit, which comprises a luciferase lysis solution, a firefly luciferase detection reagent, a NanoLuc luciferase detection buffer, a NanoLuc luciferase substrate concentrate, an AkaLuc luciferase detection buffer and an AkaLuc luciferase substrate concentrate. The firefly-NanoLuc-AkaLuc three-luciferase reporter gene detection kit prepared by the application has high detection sensitivity, a wider linear detection range, higher signals, a smaller signal attenuation degree, a slower light signal decline trend, and is more suitable for application in high-throughput detection.

A cell-based screening system and method for identifying compounds that selectively modulate the expression of CAG repeat-containing RNA associated with spinocerebellar ataxias and related disorders. The system comprises a human HEK293T cell line engineered to co-express two reporter constructs: a CAG repeat-expanded polyglutamine-nanoluciferase fusion protein with at least 60 CAG repeats, and a control firefly luciferase with no CAG repeats. Each construct contains a unique probe-binding sequence downstream of the repeat region, enabling independent quantification via multiplex RT-qPCR with fluorescent probes, as well as dual luciferase assays. The cell line is optimized for high-throughput screening to identify therapeutic compounds that reduce pathogenic CAG repeat RNA levels while sparing control transcripts. The invention further encompasses methods for screening, validating, and identifying candidate therapeutics for CAG expansion disorders, including spinocerebellar ataxias and Huntington's disease.

This invention relates to a quantitative method for ultrasonic disruption of *E. coli* cells, specifically a method based on firefly luciferase in *E. coli* cells. The method involves mixing *E. coli* expressing firefly luciferase as an internal standard with a suspension of target protein-expressing bacteria, followed by ultrasonic disruption of the cells. The degree of disruption of the target protein-expressing bacterial suspension is quantitatively calculated by measuring the activity of the firefly luciferase. The target protein-expressing bacterial suspension uses *E. coli* expressing the target protein. Compared with existing technologies, this invention overcomes the shortcomings of complex experiments, low efficiency, and unstable accuracy in existing technologies. It achieves efficient and highly accurate quantification of the degree of ultrasonic disruption of *E. coli* cells, reduces experimental complexity, and improves research and development efficiency and accuracy. This provides a scientific basis and new approach for the development of new feed protein resources and livestock breeding research, and has broad application potential.

The invention discloses a subtropical estuary benthic fish mullet chubil AHR1b gene recombinant expression plasmid and application thereof, and belongs to the technical field of environmental toxicology. The AHR1b gene recombinant expression plasmid is prepared by introducing an AHR1b gene into an expression vector. The invention also discloses a luciferase reporter gene system, which comprises an AHR1b gene recombinant expression plasmid, an ARNT1 gene recombinant expression plasmid and a firefly luciferase reporter plasmid containing a mouse CYP1A1 gene promoter. According to the invention, a luciferase reporter gene system is utilized to carry out comprehensive characterization on TCDD toxicity and evaluate the sensitivity and the specificity of a mugilogobius chuchuyi AHR1b and ARNT1 subtype dependency reporter gene system, so that a theoretical basis is provided for supporting high-throughput and high-sensitivity environment monitoring, and meanwhile, a species-specific early warning technology is provided for offshore and estuary DLCs pollution.

The invention relates to the field of bioengineering, in particular to nucleic acid, mRNA and application thereof. The invention provides nucleic acid for coding luciferase. The nucleic acid has a nucleotide sequence as shown in SEQ ID NO: 1 or SEQ ID NO: 2. According to the artificial codon optimization method provided by the invention, a plurality of version DNA sequences for highly expressing firefly luciferase are obtained by optimizing the proportion of each codon in a DNA sequence, the GC content range, the sequence repetition condition, the RNA secondary structure, the RNA free energy and the like. Compared with wild-type firefly luciferase protein, the expression quantity of other contrasts in a cell experiment is improved by 2-201 times, the fluorescent brightness is obviously improved in a mouse experiment, and the application value of firefly luciferase serving as a reporter gene is greatly improved.

The invention provides a chikungunya virus invasion model, an establishment method and application, according to the invasion model, chikungunya structural protein expression plasmids and firefly luciferase reporter gene plasmids containing specific packaging signals are co-transfected to mammalian cells, and the chikungunya structural protein expression plasmids and the firefly luciferase reporter gene plasmids containing the specific packaging signals are co-transfected by utilizing a packaging mechanism of the packaging signals. And self-assembling to form the high-simulation virus-like particle capable of efficiently bearing the reporter gene. According to the method, evaluation of chikungunya virus invading cells without depending on a BSL-3 experimental environment is achieved, and drugs / vaccines for resisting the chikungunya virus can be quantitatively screened through firefly luciferase. The method has the advantages that the whole process that viruses invade host cells can be efficiently and accurately simulated, meanwhile, high sensitivity and operation convenience are achieved, and a stable, reliable and high-drug-effect-resolution experimental platform is provided for deeply researching the invasion mechanism of CHIKV and developing targeted inhibitor high-throughput screening.

The invention discloses a firefly luciferase and NanoLuc luciferase double reporter gene detection kit. The firefly luciferase and NanoLuc luciferase double reporter gene detection kit comprises a luciferase lysis solution, a firefly luciferase detection reagent, a NanoLuc luciferase detection buffer solution and a NanoLuc luciferase substrate (50 *). Tests show that the firefly luciferase and NanoLuc luciferase double-reporter gene detection kit prepared by the invention is high in detection sensitivity, wider in detection linear range, higher in signal, small in signal attenuation degree and slower in luminescence signal descending trend, and is more suitable for being applied to high-throughput detection.

The present invention discloses a firefly luciferase-labeled Talaromyces marneffei strain and a construction method, the strain is named TM-LUC strain, and its microbial preservation number is: CGMCC No.40152; the firefly luciferase-labeled Talaromyces marneffei strain and the construction method of the present invention include: S1, constructing a firefly luciferase recombinant plasmid; S2, transferring into the genome of TM. The present invention transfers firefly luciferase into the TM genome, so that TM can encode luciferase, and bioluminescence is performed under the action of substrate luciferin, replacing the traditional ATCC18224 strain. After TM-LUC infects animals, intraperitoneal injection of luciferin substrate can theoretically make the strain glow, thereby realizing the monitoring of strain distribution in the animal body; the treatment process after administration can also be monitored; and a new direction is provided for the exploration of latent infection mechanisms.

The invention provides a biological activity detection kit based on a luciferase reporter gene. Comprising a substrate buffer solution and a firefly luciferase substrate, the substrate buffer solution comprises a zwitterionic buffer agent, glycerol, magnesium salt, a sulfydryl reducing agent, a high-energy compound, a coenzyme, a nonionic surfactant, a sulfo-nucleobase compound, an antioxidant, a metal ion masking agent and polyamine; the substrate buffer solution and a firefly luciferase substrate form a luciferase reporter gene-based biological activity detection test box; the kit not only has the advantage that one-step sample adding operation is simple, convenient and rapid, but also is not easily influenced by mixed sample adding conditions, good in result repeatability and more accurate in detection, meanwhile, the kit disclosed by the invention is brighter in light signal output and higher in sensitivity, and the detection luminescence value is higher than that of a kit of a similar product. And for some experiment systems for trace expression of firefly luciferase, the requirement of detecting the luminescence value can be met.

The application discloses a luciferase substrate, a preparation method and use thereof. The luciferase substrate has the advantages of long emission wavelength, high light intensity and high detection sensitivity after acting on firefly luciferase, and has good performance in live cell and live animal imaging, and can be used as an important analysis tool in the fields of live bio-luminescence imaging and protein quantitative detection.

The invention discloses a firefly-NanoLuc-AkaLuc three-luciferase reporter gene detection kit. The firefly-NanoLuc-AkaLuc three-luciferase reporter gene detection kit comprises a luciferase lysis solution, a firefly luciferase detection reagent, a NanoLuc luciferase detection buffer solution, a NanoLuc luciferase substrate concentrated solution, an AkaLuc luciferase detection buffer solution and an AkaLuc luciferase substrate concentrated solution. Tests show that the firefly-NanoLuc-AkaLuc triluciferase reporter gene detection kit prepared by the invention is high in detection sensitivity, wider in detection linear range, higher in signal, small in signal attenuation degree and slower in light-emitting signal descending trend, and is more suitable for being applied to high-throughput detection.

The invention discloses a method for identifying the translation activity of a 5 '-end cap analogue of mRNA (messenger Ribonucleic Acid), which is characterized in that a luciferase expression plasmid containing a 5' UTR (Untranslated Region), a firefly luciferase open reading frame, a 3 'UTR and a PolyA tail is used as a transcription system, and the translation activity of the 5'-end cap analogue is evaluated according to the activity of firefly luciferase expressed by a reporter gene. According to the method, differentiated translation activities can be identified for the 5 '-end cap analogues with different structures, a methodological basis is provided for accurate configuration of mRNA translation, and the method can also be used for high-throughput screening of the 5'-end cap analogues with intracellular translation activities.

The application discloses a uORF element for regulating chicken insulin receptor expression and application thereof, belongs to the field of molecular biology and genetic engineering, and the nucleotide sequence of the uORF element is shown in any one of SEQ ID NO. 1-2; the application comprises any one of the following: (1) regulating the expression of firefly luciferase in eukaryotic cells; the eukaryotic cells are chicken embryo fibroblasts and chicken preadipocytes; (2) being applied to genetic breeding of chickens, and the genetic breeding is breeding of meat chickens with reduced insulin resistance; (3) application in preparation of a reagent for regulating the expression of chicken insulin receptors. The application can regulate the expression level of INSR, improve insulin sensitivity, and provide material support for research on the mechanism of insulin in regulating blood glucose.

The invention discloses a construction method of a double-labeled cell based on firefly luciferase, which comprises the following steps: adding Puro, P2A and T2A sequences between a tdTomato fluorescent protein and a DNA sequence of the firefly luciferase, constructing a tdToma-P2A-Puro-T2A-fLuc sequence, designing a PCR (Polymerase Chain Reaction) primer, cloning the tdToma-P2A-Puro-T2A-fLuc sequence to a lentiviral vector in a homologous recombination manner, and transforming the lentiviral vector to a host cell, thereby obtaining the double-labeled cell based on the firefly luciferase. The method comprises the following steps: selecting bacterial colonies, carrying out amplification culture, carrying out amplification culture after sequencing is correct, extracting to obtain tdToma-P2A-Puro-T2A-fLuc expression plasmids, carrying out lentivirus packaging in HEK-293T cells by using a four-plasmid system, infecting the cells, and screening by using antibiotics to obtain the firefly luciferase-based double-labeled cells. According to the method, the red fluorescent protein tdTomato, the resistance gene puromycin and the firefly luciferase fLuc are respectively coupled through P2A and T2A elements, so that the combination of static and dynamic two-dimensional detection is realized, the change of cell activity can be efficiently and sensitively represented through the fLuc, the change of the cell quantity can be dynamically monitored in real time through the tdTomato by utilizing an IncuCyte instrument, and the qualitative and quantitative comprehensive analysis is facilitated.

The invention belongs to the field of virus detection, and discloses a novel coronavirus detection kit and application thereof in drug screening. The kit comprises a sample buffer solution, a positive control reagent, a detection reagent, a sampling label and a pipette. A modified firefly luciferase is used, the modified firefly luciferase carries the most effective site of action of 3CL at a non-critical site, and when coronavirus or host cells carrying viral components are present in a test sample, the 3CL enzyme in the sample destroys the modified firefly luciferase, breaks the enzyme into two parts, resulting in inactivation thereof. The coronavirus infection is diagnosed by measuring the specific enzyme activity of the coronavirus in a clinical sample, and the method is suitable for large-scale rapid preliminary screening and suitable for medical units such as community clinics with simple condition facilities; the positive sample provided by the invention needs to be further typed and diagnosed, such as typed and diagnosed by using a PCR (Polymerase Chain Reaction) method.

Aging is a process that the structure, functions and various physiological systems of an organism gradually decline after the organism experiences long-term physiological and metabolic activities, and is closely related to various chronic diseases and degenerative diseases. The aging process usually accompanies the phenomena of cell cycle stagnation, DNA damage, telomere shortening and the like. Existing senescence detection methods such as SA-beta-gal staining method, gene expression analysis and the like can provide certain senescence marker detection, but have limitations in the aspects of monitoring the senescence process in real time and capturing dynamic cell changes. Therefore, the development of an efficient, sensitive and real-time senescence detection technology is particularly important. The invention relates to an aging drug screening platform established on the basis of dual fluorescein plasmids, and real-time and accurate monitoring of cell molecular events in an aging process is realized by utilizing plasmids marked by two different fluoresceins and simultaneously using two fluorescently-marked genes, namely firefly luciferase and Renilla luciferase. By installing the promoter of the senescence marker gene hP21 or hGLB1 into pGL4.17 and pRL plasmids, senescence can be detected with high precision and high sensitivity, and the defects of a traditional detection method are overcome. The technology not only can reflect the cell change in the aging process in real time, but also can be widely applied to large-scale aging-related effective drug screening, and serves for obtaining drugs which can be applied to clinical treatment.

An ultrasensitive and long-lasting luminescence cascade can be used for point-of-care detection. A reaction chamber can receive and incubate a volume of a sample and a volume of magnetic tag solution before the combination is washed. Then, a working solution including d-luciferin-6-O-β-d-galactopyranoside (LUGAL), firefly luciferase (Fluc), and a luminescence buffer comprising ATP can be added to the reaction chamber. When the reaction chamber also includes tagged target components including beta galactosidase (GAL) then the GAL supplies luciferin intermediates that catalyze the cleavage of a protecting group from the LUGAL, generating luciferin, and the Fluc catalyzes the oxidation of D-luciferin in a presence of the ATP and O2, yielding oxyluciferin and luminescence. Repeated catalytic cycles increase a measurable signal of the luminescence. An optical sensor can capture an image of the luminescence and the presence of tagged target components can be determined based on an amount of luminescence in the image.

The invention belongs to the field of plant synthetic biology and plant gene expression regulation and control, and particularly relates to a vector for plant promoter dual-system quantitative characterization, a standardization method, a product and application. According to the method, a protoplast transient expression system and an agrobacterium-mediated leaf transient expression system are combined, a promoter characterization plasmid (pLIP) with a compact structure is used, a firefly luciferase module driven by a promoter to be detected and a renilla luciferase internal reference module driven by a 35S promoter are contained, and the firefly luciferase module and the renilla luciferase internal reference module are located in a T-DNA interval. Normalization and direct comparison of the activities of promoters in different systems are realized by calculating the ratio of dual luciferase and introducing an RPU (Relative Promoter Unit) method and taking a standard promoter (35S) as a reference. According to the scheme, the repeatability and cross-system compatibility of promoter activity data are remarkably improved, and the method is suitable for efficient screening and standardized characterization of regulatory elements in plant synthetic biology.

The invention discloses a firefly luciferase Fluc mutant and application of the firefly luciferase Fluc mutant, wild firefly luciferase Fluc from North America fireflies is taken as a research object, single mutation or combined mutation is carried out on a protein sequence of the firefly luciferase Fluc to obtain a mutant protein, and the mutant protein is used for preparing the firefly luciferase Fluc mutant. Compared with wild type Fluc, the heat stability and activity of the Fluc mutant protein are improved to a certain degree, compared with the wild type Fluc, the heat stability of the preferable mutant protein is improved by 13.2-14.5 DEG C, the activity is improved by 1.4-2.5 times, the yield is improved by 2.4-3.9 times, the purity of the obtained preferable mutant protein is higher than 99%, and it can be seen that the preferable mutant protein can be applied to the field of industrial production. The luciferase Fluc mutant protein provided by the invention has higher enzyme activity, better thermal stability and higher yield, widens the application conditions of luciferase Fluc, has higher practical application value, and is more suitable for large-scale production and industrial use.

The invention relates to a method for detecting the activity of a promoter FSP1 by using a dual-luciferase reporter gene, and relates to the technical field of molecular genetics and cell biology. Comprising the following steps: (1) constructing a firefly luciferase recombinant plasmid containing a promoter FSP1 sequence and a recombinant plasmid containing a sequence for expressing a mouse transcription factor Nrf2; (2) incubating the firefly luciferase recombinant plasmid containing the promoter FSP1 sequence, the recombinant plasmid containing the sequence for expressing the mouse transcription factor Nrf2 and an internal reference plasmid, and then transfecting cells; and (3) culturing the transfected cells, and breaking the cultured cells to detect luciferase activity so as to obtain the activity of the promoter FSP1. The method has the advantages of high sensitivity, high detection speed, low cost and no need of radioisotope.

The invention discloses a promoter of a larimichthys crocea interleukin 4 / 13B (IL-4 / 13B) gene. The sequence of the promoter is as shown in SEQ ID NO. 1. A recombinant plasmid pGL3-IL-4 / 13B and a control plasmid pGL3-Basic are used for respectively transfecting carp epithelioma EPC cells, a renilla luciferase reporter gene plasmid pRL-TK is used as an internal reference, and a dual-luciferase reporter gene detection system is used for analyzing the activity difference of firefly luciferase among different groups; and then, respectively using Poly (I: C), phytolectin PHA and inactivated pseudomonas protegens to stimulate EPC cells transfected with pGL3-IL-4 / 13B plasmids, and analyzing the change of the activity of firefly luciferase. The promoter sequence of the large yellow croaker IL-4 / 13B gene provided by the invention has the basic activity of the promoter, can start the transcription expression of a downstream reporter gene, and responds to the exogenous stimulation of Poly (I: C), PHA, inactivated pseudomonas proteinans and the like, so that an excellent experimental system is provided for researching the transcription regulation mechanism of the large yellow croaker IL-4 / 13B gene; the method can also be used for efficiently expressing exogenous genes or researching transgenic pseudosciaena crocea, and has theoretical significance and practical value.

The application discloses a subtropical estuary benthic fish Oxyeleotris marmoratus AHR1b gene recombination expression plasmid and application thereof, and belongs to the technical field of environmental toxicology. The AHR1b gene recombination expression plasmid is prepared by introducing the AHR1b gene into an expression vector. A luciferase reporter gene system is also disclosed, which comprises the AHR1b gene recombination expression plasmid, an ARNT1 gene recombination expression plasmid and a firefly luciferase reporter plasmid containing a mouse CYP1A1 gene promoter. The luciferase reporter gene system is used to comprehensively characterize TCDD toxicity, to evaluate the sensitivity and specificity of the Oxyeleotris marmoratus AHR1b and ARNT1 subtype-dependent reporter gene system, to provide a theoretical basis for supporting high-throughput and high-sensitivity environmental monitoring, and to provide a species-specific early warning technology for offshore and estuary DLCs pollution.

The invention belongs to the field of drug detection, and particularly discloses a method for detecting antibody-dependent cell-mediated cytotoxic biological activity of human immune globulin, which comprises the following steps: 1) adding a target cell suspension into a pore plate, respectively adding human immune globulin solutions with a series of concentrations for incubation, and finally adding an effector cell suspension for incubation; (2) detecting fluorescence of the incubated pore plate in the step (1), drawing a dose effect curve and calculating half effect concentration; the target cell is a CHO-K1 cell for expressing SARS-CoV-2 Spike protein, and the effector cell is a Jurkat cell which stably expresses CD16a on a cell membrane and is stably transfected with a firefly luciferase gene which is driven to express by an NFAT response element. The detection method disclosed by the invention can be used for accurately and reliably detecting the antibody-dependent cell-mediated cytotoxic biological activity of two different human immune globulin preparations, namely IVIG and SCIG.