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@#ObjectiveTo prepare colloidal gold immunochromatographic test paper for rapid detection of Legionella pneumophila(LP)and test its performance to ensure that it meets the national clinical diagnostic standards.MethodsLP colloidal gold immunochromatographic test paper was prepared based on double antibody sandwich ELISA,and tested for the cross reactivity,anti-interference,sensitivity,hook effect,stability and other aspects.ResultsLP colloidal gold immunochromatography test paper showed no cross reaction with 22 common pathogens in respiratory tract such as Moraxella catarrhalis,and was not affected by internal and external interferences in respiratory tract;The minimum detection limit for LP was 2.00 × 105cfu/mL,with good sensitivity and no hook effect;Under the conditions of accelerated aging at 45 ℃,simulated high temperature transportation and frozen transportation,the repeatability and stability of test paper were not affected,and the stability was good in the same batch and between different batches.ConclusionThe prepared LP colloidal gold immunochromatographic test paper realized rapid detection of LP,which was simple to operate and had good application prospect and popularization value.
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@#Objective To develop a colloidal gold immunochromatographic test strip for rapid and accurate detection of Pseudomonas aeruginosa(P.aeruginosa,Pa).Methods After bioinformatics analysis of Pa outer membrane protein OprF,the gene sequence with abundant antigenic determinants and high intraspecific homology was chemically synthesized,and then connected to pET-28a(+)vector to construct the expression vector pET-28a-OprF,which was transformed into E.coli BL21(DE3)and induced by IPTG. The recombinant OprF protein was purified by Ni Sepharose~(TM)6 Fast Flow and used to immunize two female BALB/c mice for 3～4 times by multi-point subcutaneous injection in the back at the first immunization and intraperitoneal injection at subsequent immunizations. The monoclonal antibodies were screened by animal cell fusion technique,and the colloidal gold immunochromatographic test strip for rapid detection of Pa was prepared by using monoclonal antibody and double antibody sandwich immunochromatography technique. The specificity,sensitivity and stability of the test strip were evaluated.Results Two monoclonal antibodies,Pa-1# and Pa-2#,were obtained with the titer of 1∶409 600,and both of them recognized OprF specifically. The prepared colloidal gold immunochromatographic test strip showed a sensitivity of 1. 0×10~6CFU/mL and had no cross reaction with 9 common respiratory pathogens with a good stability.Conclusion The prepared colloidal gold immunochromatographic test strip can detect Pa rapidly within 15 min,with high specificity and good stability.
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The aim of this study was to develop a semi-quantitative immunochromatographic method for rapid detection of Newcastle disease virus (NDV) antibodies by expressing HN protein in rice endosperm bioreactor. The recombinant plasmid pUC57-HN was digested by MlyⅠ and XhoⅠ to retrieve the HN gene, while the intermediate vector pMP3 containing promoter, signal peptide and terminator was digested by NaeⅠ and XhoⅠ. The HN gene and the linearized pMP3 were purified and ligated to form a recombinant plasmid pMP3-HN1. Subsequently, pMP3-HN1 and plant vector pCAMBIA1300 were digested by EcoRⅠ and Hind Ⅲ, and the HN1 gene was cloned into pCAMBIA1300. The recombinant plasmid pCAMBIA1300-HN1 was introduced into Agrobacterium tumefaciens EHA105 by electrotransformation, and the pCAMBIA1300-HN1 was transferred into rice callus by agrobacterium-mediated method. After dark culture, callus screening, differentiation, rooting and transplanting, transgenic rice seeds were obtained 4 months later. PCR identified that the HN gene has been inserted into the rice genome. SDS-PAGE and Western blotting indicated that the HN protein was successfully expressed in the positive rice endosperm. The purity of the HN protein was more than 90% by SP cation exchange chromatography and gel filtration chromatography. According to the national standards for the diagnostic techniques of Newcastle disease HI test (HI≥4log2, positive antibody reaction), a colloidal gold labeled purified HN protein was used to prepare a semi-quantitative test strip by double-antibody sandwich method for rapid detection of NDV antibody. The results showed that the test strip did not cross-react with positive sera against other viruses, and the sensitivity of the test strip reached 1:102 400 for standard positive sera of Newcastle disease. Testing of a total of 308 clinical sera showed that the compliance rate of the test strip with HI test was 97.08%, and the Kappa value was 0.942. In conclusion, high purity recombinant HN protein was obtained from rice endosperm, and a simple, rapid, highly sensitive and highly specific semi-quantitative immunochromatographic strip was developed. The test strip could be used for immune evaluation of the Newcastle disease vaccine.
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Animals , Antibodies, Viral , Chickens , HN Protein/metabolism , Newcastle Disease/prevention & control , Newcastle disease virus/metabolism , Oryza/geneticsABSTRACT
8-hydroxy-2'-deoxyguanosine (8-OHdG) is a sensitive and stable biomarker for evaluating DNA oxidative damage. A rapid and sensitive colloidal gold immunochromatographic strip was developed for 8-OHdG detection by a competitive method. The sample pad (glass cellulose film), bonding pad (glass cellulose film), nitrocellulose film and absorbent pad were pasted on the polyvinyl chloride (PVC) base plate to construct the test strip. Colloidal gold (AuNPs) was prepared by the reduction of chloroauric acid with sodium citrate. 8-OHdG antibody (Ab) was coated on the outer layer of AuNPs to form Ab@AuNPs as a probe. Bovine serum albumin (BSA) and 8-OHdG were conjugated with carbodiimide hydrochloride to prepare an artificial antigen, which was used as the coating antigen of detection line. Goat anti mouse polyclonal antibody IgG was used as the coating antibody of control line. The experimental parameters were optimized including the type of nitrocellulose membrane, the formula of loading solution, and the spraying amount of gold labeled antibody. The results showed that the appropriate nitrocellulose membrane was CN 95. The optimal loading solution included BSA (1%), Tween-20 (3%), sucrose (3%) and NaCl (0.9%). The optimal spraying amount of gold labeled antibody was 4 μL. 8-OHdG can be detected by the strip under visible light, and the level of 8-OHdG in urine can be preliminarily determined by comparing the color intensity of T line and C line. The 8-OHdG concentration in urine was further calculated by the gray value of T line and the threshold of detection was 2.55 μg/L. This colloidal gold immunochromatographic strip is simple, rapid and specific for detecting 8-OHdG in human urine to preliminarily evaluate the human status.
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Animals , Mice , 8-Hydroxy-2'-Deoxyguanosine , Antibodies, Monoclonal , Gold , Gold Colloid/chemistry , Metal Nanoparticles , Sensitivity and SpecificityABSTRACT
Objective To analyze the epidemiological characteristics of influenza in preschool children in Leshan City from 2016 to 2018, and to compare the difference between antigen colloidal gold reagent and nucleic acid detection reagent. Methods A total of 1 100 patients with suspected influenza admitted to Leshan City from January 2016 to December 2018 were selected as the research objects; the clinical data and characteristics of influenza epidemiology of the patients were collected, and the antigen colloidal gold reagent and nucleic acid detection reagent were used to detect influenza. Epidemiological characteristics; using nucleic acid detection as the standard to evaluate the diagnostic value of antigen colloidal gold reagent detection methods. Results The nucleic acid detection gold standard, antigen colloidal gold reagent method to detect influenza The sensitivity and specificity of the pathogen are both greater than 80%, and the AUC of influenza pathogen detection by the antigen colloidal gold reagent method is greater than 0.90, which has high application value . From 2016 to 2018, Leshan City was dominated by type B Y and type A H3N2; the main clinical features were cough, runny nose, and fever (P<0.05); the distribution of characteristics showed that there was no statistics on the sex of children with influenza Significantly, children less than 5 years old accounted for the highest proportion, and the proportion of children in winter and Spring Festival was significantly higher than that in summer and autumn, and the difference was statistically significant (P<0.05). Conclusion From 2016 to 2018, influenza virus infections in preschool children in Leshan City were mainly type Y and type A H3N2, and showed obvious age and seasonal characteristics. The antigenic colloidal gold reagent was used in clinical screening for children, but the test was negative. Children who are at high risk should be diagnosed with nucleic acid method as soon as possible.
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The situation of prevention of non-neonatal tetanus in China is severe. Strengthening the active immunization with tetanus toxoid vaccine (TTCV) is the key to prevent the non-neonatal tetanus. Through the detection of tetanus antibody (TAB), the immune status of individual can be determined, so as to implement the active immunization of TTCV correctly. The research on TAB detection technology is stagnant in aboard, but still in a development process in China since there is a realistic demand for TAB detection. This review collects relatively limited data of TAB detection technology in China, and summarizes the techniques such as mice toxin neutralization test (MTNT), indirect hemagglutination assay (IHA), double agar gel immune diffusion test (Rubin method), enzyme-linked immunosorbent assay (ELISA) and colloidal gold (CG), in order to provide a comprehensive basis for domestic TAB detection. The TAB detection technology in China has not yet achieved international recognition due to the lack of comparative study of domestic and international institutions and reference reagents. The special domestic situation of tetanus prevention makes the research of TAB detection technology have a certain practical significance, and rapid detection reagents such as ELISA and CG method have a certain application value in China.
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The situation of prevention of non-neonatal tetanus in China is severe. Strengthening the active immunization with tetanus toxoid vaccine (TTCV) is the key to prevent the non-neonatal tetanus. Through the detection of tetanus antibody (TAB), the immune status of individual can be determined, so as to implement the active immunization of TTCV correctly. The research on TAB detection technology is stagnant in aboard, but still in a development process in China since there is a realistic demand for TAB detection. This review collects relatively limited data of TAB detection technology in China, and summarizes the techniques such as mice toxin neutralization test (MTNT), indirect hemagglutination assay (IHA), double agar gel immune diffusion test (Rubin method), enzyme-linked immunosorbent assay (ELISA) and colloidal gold (CG), in order to provide a comprehensive basis for domestic TAB detection. The TAB detection technology in China has not yet achieved international recognition due to the lack of comparative study of domestic and international institutions and reference reagents. The special domestic situation of tetanus prevention makes the research of TAB detection technology have a certain practical significance, and rapid detection reagents such as ELISA and CG method have a certain application value in China.
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Objective: To provide data reference for the clinical diagnosis of COVID-19, promote the innovation and improvement of antibody detection technology of COVID-19, and discuss the application of antibody detection of new coronavirus by evaluating the sensitivity, specificity, compliance rate and limit of antibody detection in 10 SARS-CoV-2 antibody detection kits. Methods: According to the instructions of 10 new coronavirus antibody detection kits, the specific IgM and IgG in 74 serum samples were detect- ed, and the limit of detection of each kit was tested with 10 samples diluted by a 2-fold gradient. Results: The coincidence rate of 10 kits(product A-product J)were 89.2%(66/74), 89.2%(66/74), 86.5%(64/74), 95.9%(71/74), 52.7%(39/74), 75.7%(56/74), 86.5%(64/74), 79.7%(59/74), 50.0%(37/74), 20.3%(15/74), respectively. In the detection limit tests for the kits A-J with the 10 of 2x gradient diluted samples, the performance of kit C was the best, with the limit of detection for IgM 0.16AU/ml and the limit of detection for IgG 1.00 AU/ml. Conclusion: There are significant differences in sensitivity, specificity, compliance and limit of detection of 10 new coronavirus antibody testing reagents, and the product of high sensitivity, specificity, and compliance rates should be selected for clinical applications to ensure the accuracy of test results.
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This paper introduces a kind of immune colloidal gold detector instrument from the aspects of machinery,hardware and software.The instrument first collects one image through a CMOS sensor and then analyzes the image with image processing algorithm on Linux platform.Firstly,the instrument sets and stores the parameters separately for each test item,and then calls the saved item parameters when testing the item sample.So,the instrument can be used in a variety of fields and items.In this paper,a quantitative experimental test on C-reactive protein sample was performed,and the results indicate the coefficient of determination what denoted equal to 0.99,and the repeatability is greater than 93%.
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Algorithms , Gold Colloid , Image Processing, Computer-AssistedABSTRACT
To establish a novel colloidal gold immunochromatography assay (GICA) for rapid, sensitive and accurate detection of Haemophilus influenzae infection by using the outer membrane protein P6 as detection target. First, the linear antigen epitope located in the extracellular domain of the P6 protein (GenBank accession number: AGH02799) was predicted by bioinformatics analysis. The region (62-75 aa of the protein) with strong antigen specificity was chosen and synthesized. Two rabbits were then immunized by the polypeptides (14 aa) for production of polyclonal antibodies. Then, the recombinant P6 proteins were also obtained to produce polyclonal antibodies. Finally, based on the two antibodies, a novel colloidal GICA for detection of Haemophilus influenzae infection was established and the specificity, sensitivity, repeatability and stability of this method were evaluated. At the same time, the method was tested in clinical simulation, and the plate culture method was used to verify its accuracy. The test strip for Haemophilus influenzae infection was successfully prepared. The detection limit of the test strip was as low as 1×105 CFU/mL and the whole process can be completed within 15 minutes. The strip specifically recognized Haemophilus influenzae and did not react with nine of other common respiratory pathogens such as Streptococcus pneumoniae, Moraxella catarrhalis, Mycoplasma pneumonia, and Legionella pneumophila. And the strips could be stored at 25 °C for at least 6 months without losing sensitivity or specificity. The coincidence rate between the results of 200 clinical samples and the plate culture method was 90.5%. Haemophilus influenzae protein P6, which possessed a high degree of surface antigen accessibility and antigencity, could be used as a marker for Haemophilus influenzae detection. The immunochromatographic colloidal gold test strip which bears the features of rapidity, convenience and sensitivity provides a unique tool for the on-site surveillance and diagnosis of Haemophilus influenzae infection in clinical test.
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Animals , Humans , Rabbits , Chromatography, Affinity , Diagnostic Tests, Routine , Reference Standards , Gold Colloid , Chemistry , Haemophilus Infections , Diagnosis , Haemophilus influenzae , Limit of Detection , Sensitivity and SpecificityABSTRACT
Rice stripe virus (RSV) causes dramatic losses in rice production worldwide. In this study, two monoclonal antibodies (MAbs) 16E6 and 11C1 against RSV and a colloidal gold-based immunochromatographic strip were developed for specific, sensitive, and rapid detection of RSV in rice plant and planthopper samples. The MAb 16E6 was conjugated with colloidal gold and the MAb 11C1 was coated on the test line of the nitrocellulose membrane of the test strip. The specificity of the test strip was confirmed by a positive reaction to RSV-infected rice plants and small brown planthopper (SBPH), and negative reactions to five other rice viruses, healthy rice plants, four other vectors of five rice viruses, and non-viruliferous SBPH. Sensitivity analyses showed that the test strip could detect the virus in RSV-infected rice plant tissue crude extracts diluted to 1:20480 (w/v, g/mL), and in individual viruliferous SBPH homogenate diluted to 1:2560 (individual SPBH/µL). The validity of the developed strip was further confirmed by tests using field-collected rice and SBPH samples. This newly developed test strip is a low-cost, fast, and easy-to-use tool for on-site detection of RSV infection during field epidemiological studies and paddy field surveys, and thus can benefit decision-making for RSV management in the field.
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OBJECTIVE：To explore the feasibility of using colloidal gold immunochromatography for quantitative detection of aflatoxin B1 in traditional Chinese medicine. METHODS：Negative samples were used to investigate matrix interference by different levels of spikes.The rapid inspection performance was evaluated by examining the precision, sensitivity, linearity, repeatability and recovery rate. The sample was determined by rapid test method and verified by HPLC. RESULTS：High-concentration and low-concentration aflatoxin B1 reference materials were added to the negative sample matrix. After the measurement, it was found that there were matrix interferences in the samples such as tangerine peel and cassia seed, and the interference was greater when the concentration was increased. So high dilution factor was used to reduce the interference. The precision RSD of the rapid test method was 4.6% (n=10), the reproducibility RSD was 4.1% (n=6), and the recoveries of different samples were between 72.8% and 112.8%. The overall performance of the method was good. A total of 43 batches of 19 kinds of medicinal materials such as silkworm, cockroach and leeches were detected by two methods. The coincidence rate between the fast test and the HPLC test was 83.7%. Therefore, the results obtained by the two detection methods were considered to be approximate. CONCLUSION：Colloidal gold immunochromatographic rapid test method can be used for the quantitative detection of aflatoxin B1 in some traditional Chinese medicines, and provides technical support for the establishment and improvement of relevant rapid detection standard methods.
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@#Abstract：Objective To develop a colloidal gold immunochromatographic strip ⁃ based method for the rapid detection of Zika virus（ZIKV）NS1 antigen. Methods The gold nanoparticles modified with the anti⁃ZIKV NS1 monoclonal antibody as the detection probe were coated on the glass ⁃fiber pad. The anti ⁃ZIKV NS1 monoclonal antibody and the goat anti ⁃mouse polyclonal antibody were immobilized on a nitrocellulose membrane as the test line and the control line，respectively. In order to achieve critical results，the ratio of the optical density （OD）of the test line to that of the control line was compared. Serial diluted ZIKV NS1 standard antigen was applied to evaluate sensitivity of the immunoassay. The culture supernatant and serum samples for arboviruses（ZIKV，Dengue virus, Japanese encephalitis virus and Chikungunya virus） were utilized to demonstrate the specificity of the method. Results The detection result could read by naked eyes within 20 minutes. The visual cut ⁃off level for the test strip was achieved at 100 ng/mL of the Zika virus NS1 standard antigen. No cross⁃reactions with Dengue virus，Japanese encephalitis virus and Chikungunya virus were observed. The strip could remain good stability within 36 weeks whether stored in 4 ℃ or room temperature（22-25 ℃）. Conclusion Apart from stability， the method was convenient，rapid and specific for ZIKV NS1 antigen，which showed a promising potential in the point of care test and the screening test.
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Lateral flow assay is widely used in the point-of-care testing on-site and in-home testing with the advantage of being simple, rapid, sensitive and cost-effective. Proper labels are the key factors in lateral flow assay. Traditional labels include colloidal gold, selenium nanoparticle, and carbon nanoparticle, among which the colloidal gold is most commonly used. Lateral flow assay has been improved as a result of the discovery of new labels, such as quantum dots and nanozyme recently. Meanwhile, transformation of qualitative detection to quantitative detection is gradually realized. This article aims at introducing the most often used and the latest lateral flow assay labels, providing a basis theoretical investigation on screening proper labels for lateral flow assay researchers.
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Objective@#To evaluate the practical efficacy of a colloidal gold (CG) detection reagent of rabies virus antibody.@*Methods@#Series dilutions of rabies immunoglobulin and serum samples of rabies vaccine immunized population were tested by a CG detection reagent of rabies virus antibody and rapid fluorescent focus inhibition test (RFFIT). The consistency of the qualitative results of rabies virus antibodies between the two methods were compared. The comparison of rates was made by Chi-square test.@*Results@#For rabies immunoglobulin diluent, the detection limit of the rabies virus antibody CG detection reagent was higher than 6.53 IU/ml but lower than 9.53 IU/ml. For the serum samples, the detection limit of the rabies virus antibody CG detection reagent was higher than 2.80 IU/ml but lower than 3.30 IU/ml. The positive rates of serum rabies virus antibodies detected by CG and RFFIT were 26% and 67% respectively, and the difference was statistically significant (χ2 =13.66, P=0.000). The results of RFFIT were regarded as gold standard, false negative results but no false positive results were observed when CG was used to detect serum rabies virus antibodies.@*Conclusions@#The sensitivity of the rabies virus antibody CG detection reagent is poor, and attention should be paid to the phenomenon of missing some practically positive results in practical application of CG detection reagent.
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In this study,we estimated the application value of detecting Mycobacterium tuberculosis (MTB) specific IgG/IgM antibodies for tuberculosis diagnosis with colloidal gold immunochromatography assay (GICA).We collected 332 effective serum samples and their background information,including 260 patients with tuberculosis and 72 healthy individuals.The means of GICA was used to detect MTB specific IgG/IgM antibodies.Results were compared with the clinical diagnosis and the results of bacteriological tests.The SPSS 22.0 software was used to analyze the results,and when P＜0.05 the difference was statistically significant.The sensitivity and specificity of GICA were 41.15％ and 91.67％,and the sensitivity of the bacterial positive and negative patients were 51.38％ and 33.77％,respectively.The positive rate of IgG/IgM antibodies detection with GICA (41.15％) was much higher than that of bacteria with acid-fast stain of sputum smear (18.84％) and sputum bacteria cultivation (36.15 ％) (P ＜ 0.05) respectively.The positive rate of the combination of tuberculosis antibody detection,sputum bacterial culture and sputum smear was 61.54％,higher than the result of single method or combination of two methods.The detection of specific antibodies against MTB in serum with GICA is sensitive,specific,rapid and convenient,which can be used in clinical screening.Meanwhile,there are still certain limitations of this method,and the sensitivity and specificity need to be improved.Therefore,the GICA can be used as an auxiliary diagnosis combined with sputum bacteriology,imaging test and clinical features rather than diagnose tuberculosis alone.
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Objective To establish a rapid method for the detection of SMCY antigen. Methods To Use the technology of colloidal gold immunochromatography with double antibody sandwich assay, the gold labeled pad was coated with colloidal gold labeled SMCY rabbit polyclonal antibody, colloidal gold strip was made for detection of serum, which include 12 serum samples of pig, cattle, dog, chicken and mice and 50 serum samples of human. Results The colloidal gold test strips showed obvious specificity in human and common animal sera and could distinguish between male and female sera. Conclusion This method can be used to identify the serum of women and men, and has certain species specificity, which provides a direction for forensic science to rapidly identify gender of human samples.
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Objective To investigate the clincial value of ELISA combined with colloidal gold immunochromatography in the detection of anti-cyclic citrullinated peptide (CCP) antibody in patients with rheumatoid arthritis (RA).Methods From October 2014 to October 2016,120 patients with RA who admitted to Fuyang People 's Hospital were selected as RA group,and 120 healthy subjects were selected as the control group.Serum samples of patients were collected and tested for anti-CCP antibody by ELISA and ELISA combined with colloidal gold immunochromatography.Results ELISA method was lower than ELISA combined with colloidal gold immunochromatography test (x2 =3.943,P ＜ 0.05).The sensitivity,specificity,positive predictive value,negative predictive value of ELISA combined with colloidal gold immunochromatography assay were 93.33％,91.67％,84.85％,96.49％,respectively,which were higher than single ELISA method (all P ＜ 0.05).Conclusion ELISA combined with colloidal gold imnmunochromatography assay for detection of serum anti-C.CP antibody in patients with RA is effective and can obtain high diagnostic sensitivity and specificity,and it is worthy of promotion.
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Objective:Preparation of monoclonal antibodies against Carprofen,which lays a foundation for the rapid detection of Carprofen.Methods:Via an active ester method,Carprofen was conjugated to bovine serum albumin (BSA) as immune antigen and ovalbumin (OVA) as detection antigen,respectively.Hybridomas were obtained by fusing mouse myeloma cells SP2/0 with splenocytes from the mice immunized with Carprofen-BSA.Hybridomas 51C3 secreting antibodies against Carprofen were obtained and subcloned.Results: Ascites of monoclonal antibodies (McAb) were prepared by injecting cells of hybridoma 51C3 into mice abdomen.The titer of purified McAb was 3.2×105and the McAb was IgG1 subtype.McAb was highly specific for Carprofen and the cross-reactivity (CR50%) was less than 0.04% with Ibuprofen, Ketoprofen, Naproxen, Feinuoluofen, Indoprofen, Fenbufen respectively.The sensitivity of the McAb to carprofen was 0.32 ng/ml and the IC50value was 0.882 ng/ml.The recoveries of Carprofen in the pork samples were from 81.4% to 104.4 % and coefficients of variation were from 16.3% to 25.8%.Conclusion:All results indicate that the McAb 51C3 is suitable to develop an immunoassay colloidal gold rapid dipstick test.
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Objective Comparison the coincidence rate in the colloidal gold method and the passive agglu-tination method to detect mycoplasma pneumoniae (MP) infection, discuss the clinical value in rapid diagnosis of MP infection in the two methods. Methods Two-hundred patients with MP infection, including 100 cases in the the children group, and 100 cases in the adult group, were detected in MP-IgM antibody in serum with the colloidal gold method and the passive agglutination method. Results The positive rate of MP-IgM antibody with the passive agglutination method were slightly higher than that of the colloidal gold method in the children group (P > 0.05), While the positive rate of MP-IgM antibody with the passive agglutination method in the adult group were signifi-cantly higher than that of the colloidal gold method (P<0.05). When the antibody titer of MP-IgM antibody were 1:60, ≥1:320 in the children group, the coincidence rate of the positive results with the colloidal gold method and the passive agglutination method were 95.40%, 95.30%;When the antibody titer of MP-IgM antibody were 1:80, 1:160,≥1:320 in the adult group, the coincidence rate of the positive results with the colloidal gold method and the passive agglutination method were 0, 61.90%, 63.80%. Conclusions In the pediatric MP infection, for the high an-tibody titer of MP-IgM antibody, the positive coincidence rate with the colloidal gold method can reach clinical diag-nostic requirements. Clinical physicians according to the age and disease process of patients choose the appropriate method in order to realize the simple, rapid and accurate diagnosis of mycoplasma pneumoniae infection.