CN112964882A - Protein chip for detecting human body antimicrobial immunoglobulin and application thereof - Google Patents

Abstract

The invention discloses a protein chip for detecting human body antimicrobial immunoglobulin, wherein the immunoglobulin is IgG or IgM, the protein chip comprises a solid phase carrier and an antigen probe group for detecting the human body antimicrobial immunoglobulin, and each antigen probe of the antigen probe group is respectively fixed on the solid phase carrier in a cross-linked manner; the antigen probe set comprises two or more combinations of a probe set A, a probe set B, a probe set C, a probe set D and a probe set E. The invention also discloses a preparation method and application of the protein chip. The protein chip of the invention has the advantages of high flux, high sensitivity, good accuracy, rapidness, low cost and the like.

Description

Protein chip for detecting human body antimicrobial immunoglobulin and application thereof

Technical Field

The invention relates to the field of immunodetection, in particular to a high-flux protein chip for detecting microorganisms and a preparation method and application thereof.

Background

With the further intensive development of the research work of molecular biological chip technology, the NDA chip technology has been gradually applied to the detection and comparative study of the expression of various known or unknown nucleic acid sequences in biological samples. However, since a protein that is a functional component for performing a chemical reaction in a living cell does not show a direct relationship between a corresponding part and mRNA expressed by an active gene, the application of the cDNA chip technology as a platform for high-throughput gene expression analysis is limited to a certain extent. In addition, since various minor chemical changes in the structure and conformation of proteins can cause changes in activity or function, further studies on the functions of proteins are necessary to further reveal the relationship between various metabolic processes and proteins in cells and the molecular mechanisms of certain diseases. With the continuous maturation of DNA chip technology and the remarkable results of gene research, the development of protein function research and related technologies is further promoted, and protein chip technology is also developed.

The basic principle of protein chip technology is that various proteins are fixed on various carriers such as a titer plate, a filter membrane, a glass slide and the like in order to form a chip for detection, then the chip reacts with detected plasma or serum, anti-human IgG or IgM labeled with specific fluorescence or enzyme reacts with the chip, components which are not complementarily combined with the proteins on the chip are washed away by bleaching, then the fluorescence intensity of each point on the chip is measured by using a fluorescence scanner or laser confocal scanning technology, and the relationship of the interaction between the proteins is analyzed by the fluorescence intensity, thereby achieving the purpose of measuring the functions of various proteins.

However, in the practical application process, due to the diversity and functional complexity of proteins in biological cells, the structures of the protein probes specifically bound on the corresponding chips are different, which brings a problem that different types of detection are required, especially, a large number of samples and chips are required for detection, the detection time is greatly increased, and the cost is huge. Therefore, the development and establishment of high-throughput protein core technology with multi-sample parallel processing capability, capable of rapid analysis, would be beneficial to simplify and accelerate the progress of protein function studies. In addition, the existing chip for detecting protein still has certain defects in the aspects of sensitivity, efficiency, application scene, cost and the like.

Disclosure of Invention

The invention aims to overcome the defects in the prior art, provides a protein chip for detecting human antimicrobial immunoglobulin with high sensitivity, good accuracy, rapidness and low cost, and also provides a preparation method and application of the protein chip.

In order to achieve the purpose, the invention adopts the following technical scheme:

a protein chip for detecting human antimicrobial immunoglobulin, wherein the immunoglobulin is IgG or IgM, the protein chip comprises a solid phase carrier and an antigen probe group for detecting the human antimicrobial immunoglobulin, and each antigen probe of the antigen probe group is respectively fixed on the solid phase carrier in a crosslinking way;

the antigen probe group comprises two or more combinations of a probe group A, a probe group B, a probe group C, a probe group D and a probe group E;

the probe group A is selected from two or more than two combinations of antigen probes for detecting human immunoglobulin resisting the microorganisms shown in sequence numbers 1) to 248): 1) helicobacter pylori, 2) streptococcus pneumoniae, 3) staphylococcus saprophyticus, 4) cornflower, 5) lactobacillus gasseri, 6) gonococcus, 7) treponema pallidum, 8) erysipelothrix rubrum, 9) chlamydia psittaci, 10) brucella, 11) acinetobacter baumannii, 12) shigella dysenteriae, 13) enterococcus faecalis, 14) bacteroides ovatus, 15) burkholderia, 16) ureaplasma urealyticum, 17) neisseria meningitidis, 18) nocardia asia, 19) cryptosporidium, 20) streptococcus lipolyticus, 21) brevibacterium, 22) vibrio harveyi, 23) streptococcus agalactiae, 24) bifidobacterium dentis, 25) clostridium septicum, 26) enterobacter cloacae, 27) rickettsia rickettii, 28) bacteroides, 29) staphylococcus griseus, 30) stenotrophomonas, 31) campylobacter conjuctosus, 32) chlorella pyogenes, 33) Mycobacterium marinum, 34) Mycoplasma, 35) Vibrio cholerae, 36) Borrelia gondii, 37) Streptococcus uberis, 38) Campylobacter haemolyticus, 39) Yersinia kloni, 40) Enterobacter, 41) Clostridium cf, 42) Mycoplasma genitalium, 43) Rhodococcus, 44) Bartonella hendii, 45) Streptococcus gordonii, 46) Clostridium provenans, 47) Acinetobacter, 48) Mycobacterium tuberculosis, 49) Bordetella pertussis, 50) Myxomyces, 51) Lactococcus calophyllus, 52) Urobacter, 53) Corynebacterium corynebacterium, 54) Staphylococcus aureus, 55) Bifidobacterium animalis, 56) Neisseria, 57) Edwardsiella, 58) Streptococcus suis, 59) Tolypocladium mimicus, 60) Mycoplasma hominis, 61) Streptococcus mutans, 62) Micrococcus, 63) Clostridium caprae, 64) Streptococcus tenuis, 65) Vibrio vulnificus, 66) Bordetella, 67) Campylobacter jejuni, 68) Haemophilus parainfluenzae, 69) Rickettsia, 70) Achromobacter xylosoxidans, 71) Streptococcus micturie, 72) Salmonella enteritidis, 73) Proteus, 74) Mycobacterium columbiae, 75) Trichoderma huiensis, 76) Streptococcus monii, 77) Staphylococcus maedii, 78) Pseudomonas, 79) Legionella pneumophila, 80) Brucella suis, 81) Brucella, 82) Salmonella, 83) Sphingobacterium gallinaceum, 84) Verwensis stutzeri, 85) Staphylococcus epidermidis, 86) Clostridium spirochete, 87) Streptococcus, 88) Vibrio parahemolyticus, 89) helicobacter pylori, 90) Oribacterium sp, 91) Yersinia enterocolitica, 92) Corynebacterium pseudotuberculosis, 93) Campylobacter rectus, 94) Debardii multocida, 95) Staphylococcus aureus, 96) Mycobacterium kansasii, 97) Vibrio, 98) Acinetobacter, 99) actinomycetes, 100) Leptospira borgpetersenii, 101) Streptococcus bovis, 102) Leptochlamys oklahominis, 103) Clostridium, 104) Nocardia, 105) Afipia sp, 106) enterococcus avium, 107) Pseudomonas aeruginosa, 108) Corynebacterium diphtheriae, 109) Chlamydia pneumoniae, 110) Mycobacterium ulcerosa, 111) Bifidobacterium, 112) Escherichia albae, 113) Escherichia canis, 114) Corynebacterium caudatus, 115) Rhizobium lentum, 116) Bifidobacterium longum, 117) Markella margaria, 118) Propionibacterium, 119) Chlorella intermedia, 120) Corynebacterium urealyticum, 121) Pseudomonas putida, 122) Chlamydia trachomatis, 123) Clostridium botulinum, 124) Shigella boidinii, 125) Shigella, 126) deinocellosis, 127) Lactobacillus crispatus, 128) Lactobacillus crispatus, and, 129) Yersinia pseudotuberculosis, 130) Fusarium, 131) Bacillus thuringiensis, 132) gold, 133) Propionibacterium acnes, 134) Aeromonas hydrophila, 135) enterococcus casselii, 136) Halobacterium pseudoburkei, 137) Gardnerella vaginalis, 138) Mycobacterium septicum, 139) Burkholderia, 140) Mycobacterium volentii, 141) Yersinia pestis, 142) Aeromonas caviae, 143) Erkenella, 144) Bordetella pertussis, 145) Bordetella bronchiseptica, 146) Campylobacter Showy, 147) Klebsiella pneumoniae, 148) Streptomyces, 149) Haemophilus influenzae, 150) Anastatus actinobacillus, 151) Lactobacillus, 152) Streptococcus pyogenes, 153) Pseudomonas shigella, 154) Escherichia coli, 155) Klebsiella pneumoniae, 156) Serratia viridae, 157) Klebsiella pneumoniae, 158) Mycobacterium topterii, 159) Borrelia burgdorferi, 160) Vibrio fluvialis, 161) Serratia, 162) Klebsiella oxytoca, 163) Aeromonas, 164) Borrelia burgdorferi, 165) Morganella, 166) Bacillus pumilus, 167) Francisella tularensis, 168) Actinomyces polyfermentica, 169) Bacillus cereus, 170) Trichoderma, 1711) Clostridium difficile, 172) Vibrio alginolyticus, 173) Bacillus mucosae, 174) Acinetobacter calcoaceticus, 175) Legionella longum, 176) Clostridium perfringens, 177) Gerberia bescens, 178) Streptococcus equi, 179) Corynebacterium jejuni, 180) Citrobacter coxiella kii, 181) Proteus mirabilis, 182) Pseudomonas fluorescens, 183) Borrelia, 184) Clostridium nucleatum, 185) Corynebacterium amyloliquefaciens, 186) Klebsiella marxians, 188) Cytophilus phagocytosis, 189) Mycobacterium fraweak) Bacteroides, 190) Serratia marcescens, 191) helicobacter heptads, 192) Listeria, 193) Mycobacterium avium, 194) Clostridium tetani, 195) enterococcus kurosophora, 196) Bacillus anthracis, 197) helicobacter canadensis, 198) Lactobacillus janssensis, 199) Bordetella parapertussis, 200) Shigella flexneri, 201) Clostridium clostridia, 202) Campylobacter fetus, 203) Vibrio mimicus, 204) Brachyspira, 205) Hall's bacteria, 206) Lactobacillus, 207) Lactobacillus plantarum, 208) Corynebacterium equi, 209) Pantoea, 210) Mycobacterium abscessus, 211) M intracellulare, 212) enterococcus, 213) Clostridium difficile, 214) Legionella, 215) Klebsiella, 216) Bacillus reevesii, 217) Borrelia brueckii, 218) Treponema pallidum, 219) Bacillus, 220) Parvias monila, 221) Bifidobacterium hemolyticus, 193) Bifidobacterium, 222) Mycobacterium bovis, 223) Burkholderia dorferi, 224) Clostridium butyricum, 225) Leptospira, 226) Acinetobacter vorax, 227) Streptococcus intermedius, 228) Cronobacter, 229) Mycoplasma pneumoniae, 230) Escherichia chaffeensis, 231) Escherichia coli, 232) Enterobacter sakazakii, 233) Francisella freundii, 234) Corynebacterium ulcerosa, 235) Streptococcus sanguis, 236) Streptococcus pseudopneumoniae, 237) Actinomyces viscosus, 238) Borrelia baumannii, 239) Candida bartonia, 240) Klebsiella terreus, 241) Porphyromonas gingivalis, 242) Rickettsia typhi, 243) Haemophilus influenzae type b, 244) Streptococcus hemolyticus type A, 245) Staphylococcus albus, 246) Neisseria, 247) Bacillus diphtheriae, 248) meningococcus;

the probe group B is selected from two or more than two antigen probes for detecting human immunoglobulin resisting the microorganisms as shown in serial numbers 249) -256): 249) clostridium septicum, 250) clostridium, 251) bacteroides fragilis, 252) salmonella typhimurium, 253) escherichia coli, 254) fosetaneri, 255) streptococcus, 256) neisseria;

the probe group C is selected from two or more than two combinations of antigen probes for detecting human immunoglobulin resisting the microorganisms as shown in sequence numbers 257) -265): 257) haemophilus, 258) clostridium, 259) porphyromonas, 260) sphingomonas, 261) bacteroides, 262) haemophilus influenzae, 263) burkholderia cepacia, 264) stenotrophomonas maltophilia, 265) achromobacter;

the probe set D is selected from two or more than two antigen probes for detecting human immunoglobulin of the following microorganisms: proteobacteria, Streptococcus granulosus, Streptococcus viridans, Veillonella, Lactobacillus, Pseudomonas, Rhodococcus, Thermus, Ralstonia, Mycobacteria, TM7, Megasphaera, Actinidia, Sinomonas, Cellulomonas capnography, Streptococcus, EB virus, Mycobacterium tuberculosis, Streptococcus granulosus in oral cavity, anaemia, Legionella, Haemophilus influenzae and other gram-negative bacteria, Micrococcus, influenza B virus, Candida and Aspergillus;

the probe set E is selected from two or more than two antigen probes for detecting human immunoglobulin of the following microorganisms: JC Virus, XMRV Virus, AIDS Virus (HIV), Hepatitis C Virus (HCV), adult T cell leukemia Virus (ATLV), Herpes Simplex Virus (HSV), polyoma Virus (BK), monkey kidney Virus SV40(Simian Virus 40), Cytomegalovirus (CMV), Measles Virus (Measles Virus), Bovine Leukemia Virus (BLV), Vaccinia Virus (Vaccinia Virus), herpes Virus (HHV), herpetic stomatitis Virus (VSV), human T cell leukemia Virus (HTLV), human herpes Virus (HHV-8), Human Papilloma Virus (HPV), Seneca Valley Virus (Seneca Valley Virus), Vesicular Stomatitis Virus (VSV), small DNA Virus, mouse Virus (MMTV), EB Virus (EBV), mumps Virus, influenza A/B Virus, epidemic encephalitis B Virus, Louis encephalitis Virus, poliovirus, Varicella virus, adenovirus type 4, adenovirus type 7, HPV virus, H1N1 influenza virus, 384), H3N2 influenza virus, H5N1 influenza virus, Hepatitis A Virus (HAV), Hepatitis B Virus (HBV), Hepatitis C Virus (HCV), Hepatitis G Virus (HGV), Hepatitis D Virus (HDV), herpes simplex virus-1 (HSV-1), herpes simplex virus-2 (HSV-2), human enterovirus (human enteroviruses, HEVs), permanent beili virus (Ljungan virus), Coxsackie B virus (Coxsackie B virus), transfusion-transmitted virus (TTV), Hepatitis E Virus (HEV), varicella-zoster virus, varicella virus and hemorrhagic fever virus, brucella, forest encephalitis virus, paratyphoid bacillus, yellow fever virus.

In the present invention, it is preferable that each antigen probe of the antigen probe set is distributed in an array on the solid support.

In the present invention, it is preferable that the solid phase carrier includes a substrate and a cross-linked base material provided on a surface of the substrate, and the cross-linked base material is capable of cross-linking each antigen probe and fixing each antigen probe to the cross-linked base material.

In the present invention, a preferable scheme is that the cross-linked base material is one of a microsphere, a microporous plate and a fibrous membrane.

In the present invention, it is preferable that the substrate is a glass plate or a high molecular polymer plate, and the high molecular polymer is polyethylene, polypropylene, or nylon.

The invention also provides a preparation method of the protein chip for detecting the human body antimicrobial immunoglobulin, which comprises the following steps:

1) respectively preparing each antigen probe in the antigen probe group;

2) crosslinking each antigen probe obtained in the step 1) to a solid phase carrier to obtain the antigen probe.

In the present invention, a preferable scheme is that the step 2) specifically comprises: fixing the antigen probes obtained in the step 1) on a solid phase carrier according to array arrangement to obtain the antigen probe.

In the present invention, a preferable scheme is that the step 2) specifically comprises: modifying the surface of the solid phase carrier so that the surface of the solid phase carrier is provided with a cross-linking substrate capable of being cross-linked with the antigen probe through a covalent bond; making a dot matrix array chart, respectively dotting each antigen probe in the step 1) on each array point, fixedly connecting a corresponding antigen probe on each array point of the solid phase carrier, and drying to obtain the product.

The invention also provides a detection method, which comprises the following steps:

1) adding a serum or plasma sample to be detected on the protein chip as described in the claim 1, and reacting;

2) washing off serum or plasma samples, and then adding the secondary antibodies of the antigen probes which are marked by fluorescence or enzyme for reaction;

3) unbound secondary antibody is washed away and then detected by a fluorescence detection device.

The protein chip for detecting the human body antimicrobial immunoglobulin, which is applied to a human body antimicrobial immunocompetence analysis detection product, is disclosed; the specific detection in the application comprises the following steps:

1) adding a serum or plasma sample to be detected on the protein chip as described in the claim 1, and reacting;

2) washing off serum or plasma samples, and then adding the secondary antibodies of the antigen probes which are marked by fluorescence or enzyme for reaction;

3) unbound secondary antibody is washed away and then detected by a fluorescence detection device.

Compared with the prior art, the invention has the beneficial effects that: the protein chip has the advantages of high flux, high sensitivity, good accuracy, rapidness, low cost and the like; and the kit can be applied to a product for analyzing and detecting the human body antimicrobial immunocompetence, can effectively and quickly and accurately detect the antimicrobial immunoglobulin generated in the human body, and can further construct an analyzing and evaluating system for the human body antimicrobial immunocompetence.

Drawings

FIG. 1 is a schematic diagram of detection of Transfusion Transformed Virus (TTV) antibody by the protein chip of Experimental example 1 of the present invention;

FIG. 2 is a schematic diagram of the protein chip of Experimental example 2 of the present invention for detecting antibodies against papillomavir (HPV).

FIG. 3 is a schematic diagram of detection of TB antibodies by the protein chip of Experimental example 3 of the present invention.

FIG. 4 is a schematic diagram of detection of HBve (hepatitis B virus) antibody by the protein chip of Experimental example 4 of the present invention.

FIG. 5 is a schematic diagram of the protein chip of Experimental example 5 for detecting EBVNA1 (Epstein-Barr Virus) antibody.

Detailed Description

The present invention is further described with reference to the accompanying drawings and the detailed description, and it should be noted that any combination of the following embodiments or technical features can be used to form a new embodiment without conflict. Except as specifically noted, the materials and equipment used in this example are commercially available. The specific embodiments are merely illustrative and are not to be construed as limiting the scope of the invention.

A protein chip for detecting human antimicrobial immunoglobulin, wherein the immunoglobulin is IgG or IgM, the protein chip comprises a solid phase carrier and an antigen probe group for detecting the human antimicrobial immunoglobulin, and each antigen probe of the antigen probe group is respectively fixed on the solid phase carrier in a crosslinking way;

the antigen probe group comprises two or more combinations of a probe group A, a probe group B, a probe group C, a probe group D and a probe group E;

the probe group A is selected from two or more than two combinations of antigen probes for detecting human immunoglobulin resisting the microorganisms shown in sequence numbers 1) to 248): 1) helicobacter pylori, 2) streptococcus pneumoniae, 3) staphylococcus saprophyticus, 4) cornflower, 5) lactobacillus gasseri, 6) gonococcus, 7) treponema pallidum, 8) erysipelothrix rubrum, 9) chlamydia psittaci, 10) brucella, 11) acinetobacter baumannii, 12) shigella dysenteriae, 13) enterococcus faecalis, 14) bacteroides ovatus, 15) burkholderia, 16) ureaplasma urealyticum, 17) neisseria meningitidis, 18) nocardia asia, 19) cryptosporidium, 20) streptococcus lipolyticus, 21) brevibacterium, 22) vibrio harveyi, 23) streptococcus agalactiae, 24) bifidobacterium dentis, 25) clostridium septicum, 26) enterobacter cloacae, 27) rickettsia rickettii, 28) bacteroides, 29) staphylococcus griseus, 30) stenotrophomonas, 31) campylobacter conjuctosus, 32) chlorella pyogenes, 33) Mycobacterium marinum, 34) Mycoplasma, 35) Vibrio cholerae, 36) Borrelia gondii, 37) Streptococcus uberis, 38) Campylobacter haemolyticus, 39) Yersinia kloni, 40) Enterobacter, 41) Clostridium cf, 42) Mycoplasma genitalium, 43) Rhodococcus, 44) Bartonella hendii, 45) Streptococcus gordonii, 46) Clostridium provenans, 47) Acinetobacter, 48) Mycobacterium tuberculosis, 49) Bordetella pertussis, 50) Myxomyces, 51) Lactococcus calophyllus, 52) Urobacter, 53) Corynebacterium corynebacterium, 54) Staphylococcus aureus, 55) Bifidobacterium animalis, 56) Neisseria, 57) Edwardsiella, 58) Streptococcus suis, 59) Tolypocladium mimicus, 60) Mycoplasma hominis, 61) Streptococcus mutans, 62) Micrococcus, 63) Clostridium caprae, 64) Streptococcus tenuis, 65) Vibrio vulnificus, 66) Bordetella, 67) Campylobacter jejuni, 68) Haemophilus parainfluenzae, 69) Rickettsia, 70) Achromobacter xylosoxidans, 71) Streptococcus micturie, 72) Salmonella enteritidis, 73) Proteus, 74) Mycobacterium columbiae, 75) Trichoderma huiensis, 76) Streptococcus monii, 77) Staphylococcus maedii, 78) Pseudomonas, 79) Legionella pneumophila, 80) Brucella suis, 81) Brucella, 82) Salmonella, 83) Sphingobacterium gallinaceum, 84) Verwensis stutzeri, 85) Staphylococcus epidermidis, 86) Clostridium spirochete, 87) Streptococcus, 88) Vibrio parahemolyticus, 89) helicobacter pylori, 90) Oribacterium sp, 91) Yersinia enterocolitica, 92) Corynebacterium pseudotuberculosis, 93) Campylobacter rectus, 94) Debardii multocida, 95) Staphylococcus aureus, 96) Mycobacterium kansasii, 97) Vibrio, 98) Acinetobacter, 99) actinomycetes, 100) Leptospira borgpetersenii, 101) Streptococcus bovis, 102) Leptochlamys oklahominis, 103) Clostridium, 104) Nocardia, 105) Afipia sp, 106) enterococcus avium, 107) Pseudomonas aeruginosa, 108) Corynebacterium diphtheriae, 109) Chlamydia pneumoniae, 110) Mycobacterium ulcerosa, 111) Bifidobacterium, 112) Escherichia albae, 113) Escherichia canis, 114) Corynebacterium caudatus, 115) Rhizobium lentum, 116) Bifidobacterium longum, 117) Markella margaria, 118) Propionibacterium, 119) Chlorella intermedia, 120) Corynebacterium urealyticum, 121) Pseudomonas putida, 122) Chlamydia trachomatis, 123) Clostridium botulinum, 124) Shigella boidinii, 125) Shigella, 126) deinocellosis, 127) Lactobacillus crispatus, 128) Lactobacillus crispatus, and, 129) Yersinia pseudotuberculosis, 130) Fusarium, 131) Bacillus thuringiensis, 132) gold, 133) Propionibacterium acnes, 134) Aeromonas hydrophila, 135) enterococcus casselii, 136) Halobacterium pseudoburkei, 137) Gardnerella vaginalis, 138) Mycobacterium septicum, 139) Burkholderia, 140) Mycobacterium volentii, 141) Yersinia pestis, 142) Aeromonas caviae, 143) Erkenella, 144) Bordetella pertussis, 145) Bordetella bronchiseptica, 146) Campylobacter Showy, 147) Klebsiella pneumoniae, 148) Streptomyces, 149) Haemophilus influenzae, 150) Anastatus actinobacillus, 151) Lactobacillus, 152) Streptococcus pyogenes, 153) Pseudomonas shigella, 154) Escherichia coli, 155) Klebsiella pneumoniae, 156) Serratia viridae, 157) Klebsiella pneumoniae, 158) Mycobacterium topterii, 159) Borrelia burgdorferi, 160) Vibrio fluvialis, 161) Serratia, 162) Klebsiella oxytoca, 163) Aeromonas, 164) Borrelia burgdorferi, 165) Morganella, 166) Bacillus pumilus, 167) Francisella tularensis, 168) Actinomyces polyfermentica, 169) Bacillus cereus, 170) Trichoderma, 1711) Clostridium difficile, 172) Vibrio alginolyticus, 173) Bacillus mucosae, 174) Acinetobacter calcoaceticus, 175) Legionella longum, 176) Clostridium perfringens, 177) Gerberia bescens, 178) Streptococcus equi, 179) Corynebacterium jejuni, 180) Citrobacter coxiella kii, 181) Proteus mirabilis, 182) Pseudomonas fluorescens, 183) Borrelia, 184) Clostridium nucleatum, 185) Corynebacterium amyloliquefaciens, 186) Klebsiella marxians, 188) Cytophilus phagocytosis, 189) Mycobacterium fraweak) Bacteroides, 190) Serratia marcescens, 191) helicobacter heptads, 192) Listeria, 193) Mycobacterium avium, 194) Clostridium tetani, 195) enterococcus kurosophora, 196) Bacillus anthracis, 197) helicobacter canadensis, 198) Lactobacillus janssensis, 199) Bordetella parapertussis, 200) Shigella flexneri, 201) Clostridium clostridia, 202) Campylobacter fetus, 203) Vibrio mimicus, 204) Brachyspira, 205) Hall's bacteria, 206) Lactobacillus, 207) Lactobacillus plantarum, 208) Corynebacterium equi, 209) Pantoea, 210) Mycobacterium abscessus, 211) M intracellulare, 212) enterococcus, 213) Clostridium difficile, 214) Legionella, 215) Klebsiella, 216) Bacillus reevesii, 217) Borrelia brueckii, 218) Treponema pallidum, 219) Bacillus, 220) Parvias monila, 221) Bifidobacterium hemolyticus, 193) Bifidobacterium, 222) Mycobacterium bovis, 223) Burkholderia dorferi, 224) Clostridium butyricum, 225) Leptospira, 226) Acinetobacter vorax, 227) Streptococcus intermedius, 228) Cronobacter, 229) Mycoplasma pneumoniae, 230) Escherichia chaffeensis, 231) Escherichia coli, 232) Enterobacter sakazakii, 233) Francisella freundii, 234) Corynebacterium ulcerosa, 235) Streptococcus sanguis, 236) Streptococcus pseudopneumoniae, 237) Actinomyces viscosus, 238) Borrelia baumannii, 239) Candida bartonia, 240) Klebsiella terreus, 241) Porphyromonas gingivalis, 242) Rickettsia typhi, 243) Haemophilus influenzae type b, 244) Streptococcus hemolyticus type A, 245) Staphylococcus albus, 246) Neisseria, 247) Bacillus diphtheriae, 248) meningococcus;

the probe group B is selected from one or the combination of more than two antigen probes for detecting human immunoglobulin resisting the microorganisms as shown in serial numbers 249) -256): 249) clostridium septicum, 250) clostridium, 251) bacteroides fragilis, 252) salmonella typhimurium, 253) escherichia coli, 254) fosetaneri, 255) streptococcus, 256) neisseria;

the probe group C is selected from two or more than two combinations of antigen probes for detecting human immunoglobulin resisting the microorganisms as shown in sequence numbers 257) -265): 257) haemophilus, 258) clostridium, 259) porphyromonas, 260) sphingomonas, 261) bacteroides, 262) haemophilus influenzae, 263) burkholderia cepacia, 264) stenotrophomonas maltophilia, 265) achromobacter;

the probe set D is selected from two or more combinations of antigen probes for detecting human immunoglobulin against the microorganisms as shown in SEQ ID NO. 266) -291): 266) proteus, 267) streptococcus granulosus, 268) streptococcus viridis, 269) veillonella, 270) lactobacillus, 271) monilia, 272) rhodobacter, 273) thermus, 274) ralstonia, 275) firmicutes, 276) TM7 bacteria, 277)278) megasphaera, 279) deinsectium, 280) crescent-monomonas, 281) fibonae, 282) streptococcus, 283) EB virus, 284) mycobacterium tuberculosis, 285) fastidious, 286) legionella, 287) haemophilus influenzae, 288) micrococcus, 289) influenza b virus, 290) candida, 291) aspergillus;

the probe set E is selected from two or more than two combinations of antigen probes for detecting human immunoglobulin resisting the microorganisms as shown in sequence numbers 292) -342): 292) JC virus, 293) rabies virus, 294) XMRV virus, 295) AIDS virus, 296) adult T cell leukemia virus, 297) polyoma virus, 298) monkey kidney virus SV40, 299) cytomegalovirus, 300) measles virus, 301) bovine leukemia virus, 302) vaccinia virus, 303) herpes virus, 304) herpetic stomatitis virus, 305) human herpes virus HHV-8, 306) human papilloma virus, 307) Selenekagana virus, 308) vesicular stomatitis virus, 309) miniDNA virus, 310) mouse virus, 311) mumps virus, 312) influenza B virus, 313) epidemic encephalitis B virus, 314) St 323) Hepatitis c virus, 324) hepatitis G virus, 325) hepatitis d virus, 326) herpes simplex virus type 1, 327) herpes simplex virus type 2, 328) HEVs virus, 329) influenza a virus, 330) permanent beili virus, 331) coxsackie B virus, 332) transfusion transmitted virus, 333) hepatitis e virus, 334) varicella-zoster virus, 335) EB virus, 336) poliovirus, 337) rotavirus, 338) hemorrhagic fever virus, 339) brucella, 340) forest encephalitis virus, 341) salmonella paratyphi, 342) yellow fever virus.

The human antimicrobial immunoglobulin to be detected can be IgG antibody or IgM antibody, but generally, the content of the IgG antibody in the human body is higher, and the detection is more sensitive. In order to facilitate the preparation and detection of the protein chip, the probes forming the protein probe group are distributed on the solid phase carrier in an array manner, so that matched sample application equipment and detection equipment are more conveniently utilized.

In the present invention, it is preferable that the solid phase carrier includes a substrate and a cross-linked base material provided on a surface of the substrate, and the cross-linked base material is capable of cross-linking each antigen probe and fixing each antigen probe to the cross-linked base material.

In the present invention, a preferable scheme is that the cross-linked base material is one of a microsphere, a microporous plate and a fibrous membrane.

In the present invention, it is preferable that the substrate is a glass plate or a high molecular polymer plate, and the high molecular polymer is polyethylene, polypropylene, or nylon.

The invention also provides a preparation method of the protein chip for detecting the human body antimicrobial immunoglobulin, which comprises the following steps:

1) respectively preparing each antigen probe in the antigen probe group;

2) crosslinking each antigen probe obtained in the step 1) to a solid phase carrier to obtain the antigen probe.

In the present invention, a preferable scheme is that the step 2) specifically comprises: fixing the antigen probes obtained in the step 1) on a solid phase carrier according to array arrangement to obtain the antigen probe.

In the present invention, a preferable scheme is that the step 2) specifically comprises: modifying the surface of the solid phase carrier to enable the surface of the solid phase carrier to be provided with a cross-linking substrate (for example, the cross-linking substrate can be made of microspheres, micro-porous plates and fibrous membranes, and the cross-linking substrate is provided with immobilized protein, antibody or polypeptide which can be cross-linked with the antigen probe); making a dot matrix array chart, respectively dotting each antigen probe in the step 1) on each array point, fixedly connecting a corresponding antigen probe on each array point of the solid phase carrier, and drying to obtain the product.

The invention also provides a detection method, which comprises the following steps:

1) adding a serum or plasma sample to be detected on the protein chip as described in the claim 1, and reacting;

2) washing off serum or plasma samples, and then adding the secondary antibodies of the antigen probes which are marked by fluorescence or enzyme for reaction;

3) unbound secondary antibody is washed away and then detected by a fluorescence detection device.

The protein chip for detecting the human antimicrobial immunoglobulin, which is applied to the analysis and detection product of the human antimicrobial immunocompetence, is disclosed.

Example 1

A protein chip for detecting human body antimicrobial immunoglobulin, wherein the immunoglobulin is IgG, the protein chip comprises a solid phase carrier and an antigen probe group for detecting the human body antimicrobial immunoglobulin, and each antigen probe of the antigen probe group is respectively fixed on the solid phase carrier in a crosslinking way;

the antigen probe group comprises a probe group A, a probe group B, a probe group C, a probe group D and a probe group E;

the probe group A is selected from the combination of antigen probes for detecting human immunoglobulin resisting the microorganisms shown in the sequence numbers 1) to 248); the probe group B is selected from a combination of antigen probes for detecting human immunoglobulin against the microorganisms as shown in the above serial numbers 249) -256); the probe group C is selected from the combination of antigen probes for detecting human immunoglobulin against the microorganisms as shown in the above serial numbers 257) -265); the probe set D is selected from a combination of antigen probes for detecting human immunoglobulin against the microorganisms as shown in the above serial numbers 266) -291); the probe set E is selected from a combination of antigen probes for detecting human immunoglobulin against the microorganisms represented by the above serial numbers 292) -342).

Example 1 transfusion-transmitted Virus (TTV) detection

Referring to fig. 1, 6 positive samples and 24 negative samples of the defined transfusioned cultured virus were detected, and the detection rate of positive detection and the negative accuracy rate of the protein chip of the invention example 1 were 100%. (OD. gtoreq.0.4 positive, OD. gtoreq.0.1 negative).

Experimental example 2 detection of Human Papilloma Virus (HPV)

Referring to fig. 2, the positive rate of detection is 100% and the negative accuracy is 100% by detecting the positive samples 10 and 20 positive samples of papillomavir, which are well-defined, on the protein chip of the embodiment 1 of the present invention. (OD ≧ 0.8positive, OD ≦ 0.3 negative).

EXAMPLE 3 detection of Mycobacterium Tuberculosis (TB)

Referring to fig. 3, 15 positive samples inoculated with bcg were detected, 15 negative samples were detected, and the detection rate of positive detection was 100% and the negative accuracy was 100% by the protein chip of example 1 of the present invention. (OD ≧ 0.8positive, OD ≦ 0.1 negative).

EXAMPLE 4 detection of Hepatitis B Virus (HBV)

Referring to fig. 4, 15 positive samples and 15 negative samples infected with HBV were detected, and the detection of the protein chip of example 1 of the present invention found that the detection rate of positive was 100% and the negative accuracy was 100%. (OD ≧ 1.0positive, OD ≦ 0.1 negative).

Experimental example 5-Epstein-Barr Virus (EB) detection

Referring to fig. 5, 15 positive samples infected with EBV and 15 negative samples were detected, and the detection of the protein chip of example 1 of the present invention found that the detection rate of positive was 100% and the negative accuracy was 100%. (OD ≧ 1.0positive, OD ≦ 0.1 negative).

Experimental example 6

Mixing the transfusion transmission virus positive sample, the human papilloma virus positive sample, the mycobacterium tuberculosis positive sample, the hepatitis B virus positive sample and the EB virus positive sample of the experimental examples 1-5, and then detecting the new protein product of the embodiment 1 of the invention; mixing the negative samples of the 5 viruses, and detecting the negative samples by the novel protein product of the embodiment 1; the 5 virus detection results show that the 5 virus detection results in the positive sample are all positive, and the negative results are all negative; therefore, the protein chip of the invention has good sensitivity and detection efficiency.

Example 2

A protein chip for detecting human body antimicrobial immunoglobulin, wherein the immunoglobulin is IgG, the protein chip comprises a solid phase carrier and an antigen probe group for detecting the human body antimicrobial immunoglobulin, and each antigen probe of the antigen probe group is respectively fixed on the solid phase carrier in a crosslinking way;

the antigen probe group comprises a probe group A, a probe group B, a probe group C, a probe group D and a probe group E;

the probe set A comprises a combination of antigen probes for detecting human immunoglobulin against the following microorganisms: leptospira, chlamydia psittaci, staphylococcus aureus, bacillus anthracis, meningococcus, clostridium tetani, haemophilus influenzae type b;

the probe set B comprises a combination of antigen probes for detecting human immunoglobulin against the following microorganisms: salmonella typhimurium, escherichia coli;

the probe set C comprises a combination of antigen probes for detecting human immunoglobulin against the following microorganisms: haemophilus influenzae, achromobacter;

the probe set D comprises a combination of antigen probes for detecting human immunoglobulin against the following microorganisms: EB virus, Mycobacterium tuberculosis;

the probe group E comprises a combination of antigen probes for detecting human immunoglobulin against the following microorganisms: rabies virus, hepatitis B virus, varicella virus, hepatitis A virus, H5N1 influenza virus, hemorrhagic fever virus, human papilloma virus; brucellosis, forest encephalitis virus, paratyphoid bacillus, yellow fever virus, poliovirus, hepatitis G virus;

the protein chip is prepared by the following steps:

1) respectively preparing each antigen probe in the antigen probe group;

2) arranging and cross-linking each antigen probe obtained in the step 1) according to a 5 x 5 array on a solid phase carrier, and obtaining the antigen probe, wherein the specific arrangement is as follows: leptospira (1 × 1), staphylococcus aureus (1 × 2), bacillus anthracis (1 × 3), meningococcus (1 × 4), clostridium tetani (1 × 5), clostridium septicum (2 × 1), salmonella typhimurium (2 × 2), escherichia coli (2 × 3), haemophilus influenzae type b (2 × 4), epstein barr virus (2 × 5), hemorrhagic fever virus (3 × 1), human papilloma virus (3 × 2), brucella (3 × 3), haemophilus influenzae (3 × 4), forest encephalitis virus (3 × 5), paratyphoid bacillus (4 × 1), yellow fever virus (4 × 2), poliovirus (4 × 3), hepatitis G virus (4 × 4), mycobacterium tuberculosis (4 × 5), rabies virus (5 × 1), hepatitis b virus (5 × 2), varicella virus (5 × 3), Hepatitis A virus (5X 4), H5N1 influenza virus (5X 5).

Experimental example 7

Serum samples of 10 volunteers were taken from each volunteer, and then detected by the protein chip of the invention in example 2, and detected by fluorescence immunoassay, and the specific experimental data are as follows:

note: in the above table, a circle indicates a negative detection result, and ● indicates a positive detection result.

As can be seen from the above table, the protein chip of the embodiment 2 of the present invention can simultaneously detect a plurality of viruses with large quantity; the method can judge the capability of the human body for antimicrobial immunity through the types of the antimicrobial immunoglobulin existing in the human body by combining the condition and the medical history of the volunteers who are vaccinated once and corresponding detection data to detect and evaluate; therefore, the protein chip can be applied to products for analyzing and detecting the human body antimicrobial immunocompetence, so that the human body immunocompetence can be effectively analyzed and evaluated, and the application scene of the protein chip is expanded.

Finally, it should be noted that: the above embodiments are only preferred embodiments of the present invention, and the protection scope of the present invention should not be limited thereby, and any insubstantial changes and substitutions made by those skilled in the art based on the present invention are within the protection scope of the present invention.

Claims (10)

1. A protein chip for detecting human body antimicrobial immunoglobulin is characterized in that the immunoglobulin is IgG or IgM, the protein chip comprises a solid phase carrier and an antigen probe group for detecting the human body antimicrobial immunoglobulin, and each antigen probe of the antigen probe group is respectively fixed on the solid phase carrier in a cross-linking way;

the antigen probe group comprises two or more combinations of a probe group A, a probe group B, a probe group C, a probe group D and a probe group E;

the probe group A is selected from two or more than two combinations of antigen probes for detecting human immunoglobulin resisting the microorganisms shown in sequence numbers 1) to 248): 1) helicobacter pylori, 2) streptococcus pneumoniae, 3) staphylococcus saprophyticus, 4) cornflower, 5) lactobacillus gasseri, 6) gonococcus, 7) treponema pallidum, 8) erysipelothrix rubrum, 9) chlamydia psittaci, 10) brucella, 11) acinetobacter baumannii, 12) shigella dysenteriae, 13) enterococcus faecalis, 14) bacteroides ovatus, 15) burkholderia, 16) ureaplasma urealyticum, 17) neisseria meningitidis, 18) nocardia asia, 19) cryptosporidium, 20) streptococcus lipolyticus, 21) brevibacterium, 22) vibrio harveyi, 23) streptococcus agalactiae, 24) bifidobacterium dentis, 25) clostridium septicum, 26) enterobacter cloacae, 27) rickettsia rickettii, 28) bacteroides, 29) staphylococcus griseus, 30) stenotrophomonas, 31) campylobacter conjuctosus, 32) chlorella pyogenes, 33) Mycobacterium marinum, 34) Mycoplasma, 35) Vibrio cholerae, 36) Borrelia gondii, 37) Streptococcus uberis, 38) Campylobacter haemolyticus, 39) Yersinia kloni, 40) Enterobacter, 41) Clostridium cf, 42) Mycoplasma genitalium, 43) Rhodococcus, 44) Bartonella hendii, 45) Streptococcus gordonii, 46) Clostridium provenans, 47) Acinetobacter, 48) Mycobacterium tuberculosis, 49) Bordetella pertussis, 50) Myxomyces, 51) Lactococcus calophyllus, 52) Urobacter, 53) Corynebacterium corynebacterium, 54) Staphylococcus aureus, 55) Bifidobacterium animalis, 56) Neisseria, 57) Edwardsiella, 58) Streptococcus suis, 59) Tolypocladium mimicus, 60) Mycoplasma hominis, 61) Streptococcus mutans, 62) Micrococcus, 63) Clostridium caprae, 64) Streptococcus tenuis, 65) Vibrio vulnificus, 66) Bordetella, 67) Campylobacter jejuni, 68) Haemophilus parainfluenzae, 69) Rickettsia, 70) Achromobacter xylosoxidans, 71) Streptococcus micturie, 72) Salmonella enteritidis, 73) Proteus, 74) Mycobacterium columbiae, 75) Trichoderma huiensis, 76) Streptococcus monii, 77) Staphylococcus maedii, 78) Pseudomonas, 79) Legionella pneumophila, 80) Brucella suis, 81) Brucella, 82) Salmonella, 83) Sphingobacterium gallinaceum, 84) Verwensis stutzeri, 85) Staphylococcus epidermidis, 86) Clostridium spirochete, 87) Streptococcus, 88) Vibrio parahemolyticus, 89) helicobacter pylori, 90) Oribacterium sp, 91) Yersinia enterocolitica, 92) Corynebacterium pseudotuberculosis, 93) Campylobacter rectus, 94) Debardii multocida, 95) Staphylococcus aureus, 96) Mycobacterium kansasii, 97) Vibrio, 98) Acinetobacter, 99) actinomycetes, 100) Leptospira borgpetersenii, 101) Streptococcus bovis, 102) Leptochlamys oklahominis, 103) Clostridium, 104) Nocardia, 105) Afipia sp, 106) enterococcus avium, 107) Pseudomonas aeruginosa, 108) Corynebacterium diphtheriae, 109) Chlamydia pneumoniae, 110) Mycobacterium ulcerosa, 111) Bifidobacterium, 112) Escherichia albae, 113) Escherichia canis, 114) Corynebacterium caudatus, 115) Rhizobium lentum, 116) Bifidobacterium longum, 117) Markella margaria, 118) Propionibacterium, 119) Chlorella intermedia, 120) Corynebacterium urealyticum, 121) Pseudomonas putida, 122) Chlamydia trachomatis, 123) Clostridium botulinum, 124) Shigella boidinii, 125) Shigella, 126) deinocellosis, 127) Lactobacillus crispatus, 128) Lactobacillus crispatus, and, 129) Yersinia pseudotuberculosis, 130) Fusarium, 131) Bacillus thuringiensis, 132) gold, 133) Propionibacterium acnes, 134) Aeromonas hydrophila, 135) enterococcus casselii, 136) Halobacterium pseudoburkei, 137) Gardnerella vaginalis, 138) Mycobacterium septicum, 139) Burkholderia, 140) Mycobacterium volentii, 141) Yersinia pestis, 142) Aeromonas caviae, 143) Erkenella, 144) Bordetella pertussis, 145) Bordetella bronchiseptica, 146) Campylobacter Showy, 147) Klebsiella pneumoniae, 148) Streptomyces, 149) Haemophilus influenzae, 150) Anastatus actinobacillus, 151) Lactobacillus, 152) Streptococcus pyogenes, 153) Pseudomonas shigella, 154) Escherichia coli, 155) Klebsiella pneumoniae, 156) Serratia viridae, 157) Klebsiella pneumoniae, 158) Mycobacterium topterii, 159) Borrelia burgdorferi, 160) Vibrio fluvialis, 161) Serratia, 162) Klebsiella oxytoca, 163) Aeromonas, 164) Borrelia burgdorferi, 165) Morganella, 166) Bacillus pumilus, 167) Francisella tularensis, 168) Actinomyces polyfermentica, 169) Bacillus cereus, 170) Trichoderma, 1711) Clostridium difficile, 172) Vibrio alginolyticus, 173) Bacillus mucosae, 174) Acinetobacter calcoaceticus, 175) Legionella longum, 176) Clostridium perfringens, 177) Gerberia bescens, 178) Streptococcus equi, 179) Corynebacterium jejuni, 180) Citrobacter coxiella kii, 181) Proteus mirabilis, 182) Pseudomonas fluorescens, 183) Borrelia, 184) Clostridium nucleatum, 185) Corynebacterium amyloliquefaciens, 186) Klebsiella marxians, 188) Cytophilus phagocytosis, 189) Mycobacterium fraweak) Bacteroides, 190) Serratia marcescens, 191) helicobacter heptads, 192) Listeria, 193) Mycobacterium avium, 194) Clostridium tetani, 195) enterococcus kurosophora, 196) Bacillus anthracis, 197) helicobacter canadensis, 198) Lactobacillus janssensis, 199) Bordetella parapertussis, 200) Shigella flexneri, 201) Clostridium clostridia, 202) Campylobacter fetus, 203) Vibrio mimicus, 204) Brachyspira, 205) Hall's bacteria, 206) Lactobacillus, 207) Lactobacillus plantarum, 208) Corynebacterium equi, 209) Pantoea, 210) Mycobacterium abscessus, 211) M intracellulare, 212) enterococcus, 213) Clostridium difficile, 214) Legionella, 215) Klebsiella, 216) Bacillus reevesii, 217) Borrelia brueckii, 218) Treponema pallidum, 219) Bacillus, 220) Parvias monila, 221) Bifidobacterium hemolyticus, 193) Bifidobacterium, 222) Mycobacterium bovis, 223) Burkholderia dorferi, 224) Clostridium butyricum, 225) Leptospira, 226) Acinetobacter vorax, 227) Streptococcus intermedius, 228) Cronobacter, 229) Mycoplasma pneumoniae, 230) Escherichia chaffeensis, 231) Escherichia coli, 232) Enterobacter sakazakii, 233) Francisella freundii, 234) Corynebacterium ulcerosa, 235) Streptococcus sanguis, 236) Streptococcus pseudopneumoniae, 237) Actinomyces viscosus, 238) Borrelia baumannii, 239) Candida bartonia, 240) Klebsiella terreus, 241) Porphyromonas gingivalis, 242) Rickettsia typhi, 243) Haemophilus influenzae type b, 244) Streptococcus hemolyticus type A, 245) Staphylococcus albus, 246) Neisseria, 247) Bacillus diphtheriae, 248) meningococcus;

the probe group B is selected from two or more than two antigen probes for detecting human immunoglobulin resisting the microorganisms as shown in serial numbers 249) -256): 249) clostridium septicum, 250) clostridium, 251) bacteroides fragilis, 252) salmonella typhimurium, 253) escherichia coli, 254) fosetaneri, 255) streptococcus, 256) neisseria;

the probe group C is selected from two or more than two combinations of antigen probes for detecting human immunoglobulin resisting the microorganisms as shown in sequence numbers 257) -265): 257) haemophilus, 258) clostridium, 259) porphyromonas, 260) sphingomonas, 261) bacteroides, 262) haemophilus influenzae, 263) burkholderia cepacia, 264) stenotrophomonas maltophilia, 265) achromobacter;

the probe set D is selected from two or more combinations of antigen probes for detecting human immunoglobulin against the microorganisms as shown in SEQ ID NO. 266) -291): 266) proteus, 267) streptococcus granulosus, 268) streptococcus viridis, 269) veillonella, 270) lactobacillus, 271) monilia, 272) rhodobacter, 273) thermus, 274) ralstonia, 275) firmicutes, 276) TM7 bacteria, 277)278) megasphaera, 279) deinsectium, 280) crescent-monomonas, 281) fibonae, 282) streptococcus, 283) EB virus, 284) mycobacterium tuberculosis, 285) fastidious, 286) legionella, 287) haemophilus influenzae, 288) micrococcus, 289) influenza b virus, 290) candida, 291) aspergillus;

the probe set E is selected from two or more than two combinations of antigen probes for detecting human immunoglobulin resisting the microorganisms as shown in sequence numbers 292) -342): 292) JC virus, 293) rabies virus, 294) XMRV virus, 295) AIDS virus, 296) adult T cell leukemia virus, 297) polyoma virus, 298) monkey kidney virus SV40, 299) cytomegalovirus, 300) measles virus, 301) bovine leukemia virus, 302) vaccinia virus, 303) herpes virus, 304) herpetic stomatitis virus, 305) human herpes virus HHV-8, 306) human papilloma virus, 307) Selenekagana virus, 308) vesicular stomatitis virus, 309) miniDNA virus, 310) mouse virus, 311) mumps virus, 312) influenza B virus, 313) epidemic encephalitis B virus, 314) St 323) Hepatitis c virus, 324) hepatitis G virus, 325) hepatitis d virus, 326) herpes simplex virus type 1, 327) herpes simplex virus type 2, 328) HEVs virus, 329) influenza a virus, 330) permanent beili virus, 331) coxsackie B virus, 332) transfusion transmitted virus, 333) hepatitis e virus, 334) varicella-zoster virus, 335) EB virus, 336) poliovirus, 337) rotavirus, 338) hemorrhagic fever virus, 339) brucella, 340) forest encephalitis virus, 341) salmonella paratyphi, 342) yellow fever virus.

2. The protein chip for detecting human antimicrobial immunoglobulin of claim 1, wherein each antigen probe of the antigen probe set is distributed on the solid support in an array.

3. The protein chip for detecting human antimicrobial immunoglobulin of claim 1, wherein the solid phase carrier comprises a substrate and a cross-linking substrate disposed on the surface of the substrate, the cross-linking substrate is capable of cross-linking each antigen probe and fixing each antigen probe on the cross-linking substrate.

4. The protein chip for detecting human antimicrobial immunoglobulin according to claim 3, wherein the cross-linked substrate is one of a microsphere, a microplate and a fibrous membrane.

5. The protein chip for detecting human antimicrobial immunoglobulin of claim 3, wherein the substrate is a glass plate or a high molecular polymer plate, and the high molecular polymer is polyethylene, polypropylene or nylon.

6. The protein chip for detecting human antimicrobial immunoglobulin according to claim 1, wherein the protein chip is prepared by the following steps:

1) respectively preparing each antigen probe in the antigen probe group;

2) crosslinking each antigen probe obtained in the step 1) to a solid phase carrier to obtain the antigen probe.

7. The protein chip for detecting human antimicrobial immunoglobulin according to claim 6, wherein the step 2) is specifically: fixing the antigen probes obtained in the step 1) on a solid phase carrier according to array arrangement to obtain the antigen probe.

8. The protein chip for detecting human antimicrobial immunoglobulin according to claim 6, wherein said step 2) is specifically: modifying the surface of the solid phase carrier so that the surface of the solid phase carrier is provided with a cross-linking substrate capable of being cross-linked with the antigen probe through a covalent bond; making a dot matrix array chart, respectively dotting each antigen probe in the step 1) on each array point, fixedly connecting a corresponding antigen probe on each array point of the solid phase carrier, and drying to obtain the product.

9. The protein chip for detecting human antimicrobial immunoglobulin according to claim 1, which is used in human antimicrobial immunocompetence analysis detection products.

10. The protein chip for detecting human body antimicrobial immunoglobulin according to claim 1, which is used in a human body antimicrobial immunocompetence analysis detection product, wherein the specific detection in the application comprises the following steps:

1) adding a serum or plasma sample to be detected on the protein chip as described in the claim 1, and reacting;

2) washing off serum or plasma samples, and then adding the secondary antibodies of the antigen probes which are marked by fluorescence or enzyme for reaction;

3) unbound secondary antibody is washed away and then detected by a fluorescence detection device.

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