37 results about "Anaplasma phagocytophila" patented technology

The invention provides compositions and methods for the detection and quantification of A. phagocytophilum (formerly known as Ehrlichia equi) antibodies and antibody fragments.

The invention relates to preparation and application of a rickettsia discrimination detection gene chip. The preparation method comprises the following steps: preparing universal and specific primers, preparing seven rickettsia nucleic acid typing probes, preparing an oligonucleotide chip, establishing a multiplex PCR (polymerase chain reaction) system, and establishing a hybridization system. The gene chip can simultaneously discriminate seven important rickettsiae, including rickettsia prowazeki, rickettsia mooseri, spotted fever group rickettsia, Coxiella burnetii, Orientia tsutsugamushi, Ehrlichia chaffeensis and Anaplasma phagocytophilum. The gene chip has the advantages of high speed, high accuracy, high flux and high specificity, and can provide a novel detection means for clinical diagnosis and epidemiological investigation of rickettsia infection.

The invention provides methods and compositions for the detection and treatment of Anaplasma phagocytophilum and Anaplasma platys infection.

We have successfully sequenced and cloned the gene expressing the Major Surface Protein 5 (MSP5) of A. phagocytophilum. The recombinant MSP5 (rMSP5) protein has been tested using sera from humans and dogs infected with A. phacytophilum. The polypeptide has been found to be immunogenic and useful as a diagnostic test antigen. The polypeptide antigen of the subject invention can provide the basis of a diagnostic assay that would allow the rapid, in-house, laboratory diagnosis of infection with A. phagocytophilum using a sample (e.g., serum, plasma, or whole blood) from an infected human or animal. Additionally, the subject invention provides methods of detecting the presence of A. phagocytophilum in biological or environmental samples utilizing antibodies provided by the subject invention. Furthermore, the use of the single antigen in the diagnosis of this important disease offers many advantages including enhanced test specificity, ease of testing and consistency of results using synthetically produced test antigens instead of cultured, whole organisms.

The invention provides methods and compositions for the detection and treatment of Anaplasma phagocytophilum and Anaplasma platys infection.

Disclosed are cloning and expression of a plurality of protein fragments of virB10, a Type IV Secretion System (TIVSS) in Anaplasma phagocytophilum. Such recombinant protein fragments are useful in the ELISA detection of anaplasma pathogen. The use of same as kits for ELISA is also disclosed.

Disclosed are two (2) proteins in the Type IV Secretion System (TIVSS) in Anaplasma phagocytophilum (namely, virB 10 and virB11) useful in the ELISA detection of Anaplasma pathogen. The recombinant expression of virB 10 and virB 11 and their use as kits for ELISA are also disclosed.

The invention discloses a kit and method for detecting anaplasmosis and rickettsioses based on LAMP. The invention relates to a molecular biological detection technology for anaplasmosis and rickettsioses, provides specific primers which are designed to be directed to ompB gene of spotted fever group rickettsia (SFGR) and msp2 gene of anaplasma phagocytophilum, and provides a detection kit for SFGR and anaplasma phagocytophilum by using LAMP. The kit has the characteristics of rapidness, sensitivity, high specificity, no experimental condition restrictions, etc., and is applicable to clinicaldetection for spotted fever and granulocytic anaplasmosis, especially in rural areas where medical conditions are limited. The invention also discloses a method for detecting rickettsioses with the kit.

The invention relates to compositions and methods for the detection of various infectious organisms, including heartworm (Dirofilaria immitis), Ehrlichia Canis, Anaplasma phagocytophilum, and Borrelia burgdorferi. More particularly, this invention relates to antibodies that bind to a heartworm antigen, the E. Canis gp36 polypeptide, the A. phagocytophilum p44 polypeptide, the B. burgdorferi OspA, OspC, OspF, p39, p41 and VlsE polypeptides, and uses thereof.

Disclosed is the cloning, expression and purification of a hemolysin protein and its protein fragments in Anaplasma phagocytophilum. The recombinant hemolysin and its protein fragments are useful in the ELISA detection of anaplasma pathogen. The use of same as a kit for ELISA is also disclosed.

The invention discloses a method for detecting anaplasma phagocytophilum employing a loop-mediated isothermal amplification (LAMP) technology. The detection method comprises an LAMP primer design, building of an LAMP reaction system and adoption of an LAMP detection method. Specificity detection, sensitivity detection and fluorescent visual detection are adopted in the LAMP detection method. The anaplasma phagocytophilum 16SrRNA is taken as a target gene locus, and has the characteristics of being fast, sensitive, efficient, low in cost and the like.

Anaplasma phagocytophilum surface protein AipA and / or fragments thereof which comprise an invasin domain are used in compositions suitable to elicit an immune response to treat or prevent infections caused by tick-born bacteria of the Anaplasmatacaea family. AipA proteins and protein fragments or antibodies directed to AipA proteins and protein fragments are also used in diagnostic assays to detect exposure to and / or infection with Anaplasmatacaea. AipA and / or fragments thereof are also used for these purposes in combination with one or both of Asp14 and OmpA proteins and / or fragments thereof which comprise an invasin domain. Homologs of these proteins are also used in the compositions and assays.

Anaplasma phagocytophilum surface protein AipA and / or fragments thereof which comprise an invasin domain are used in compositions suitable to elicit an immune response to treat or prevent infections caused by tick-born bacteria of the Anaplasmatacaea family. AipA proteins and protein fragments or antibodies directed to AipA proteins and protein fragments are also used in diagnostic assays to detect exposure to and / or infection with Anaplasmatacaea. AipA and / or fragments thereof are also used for these purposes in combination with one or both of Asp14 and OmpA proteins and / or fragments thereof which comprise an invasin domain. Homologs of these proteins are also used in the compositions and assays.

The invention provides methods and compositions for the detection and treatment of Anaplasma phagocytophilum and Anaplasma platys infection.

The invention discloses an anaplasma phagocytophilum P44 recombinant protein, and belongs to the field of animal virus antibody detection. The recombinant protein comprises an amino acid sequence shown in SEQ ID NO. 1 or is composed of the amino acid sequence shown in SEQ ID NO. 1. The invention further discloses a gene of the recombinant protein, a vector and a host cell containing the gene, a preparation method of the recombinant protein and application of the recombinant protein in detection of an anaplasma phagocytophilum antibody. The recombinant protein is used for detecting the anaplasma phagocytophilum antibody, is convenient and fast to use, high in sensitivity and high in specificity, has no cross reaction with other pathogens, and has great clinical significance and wide application prospects.

The invention discloses a dual fluorescence PCR detection kit, primer and probe for simultaneously detecting anaplasma phagocytophilum and Ehrlichia chaffeensis. The detection primer and probe are shown as SEQ ID NO:1 to SEQ ID NO:6 in a sequence table, wherein the sequences SEQ ID NO:1 and SEQ ID NO:2 respectively refer to a forward primer and a reverse primer for detecting the anaplasma phagocytophilum; the sequence SEQ ID NO:3 refers to a fluorescence probe for detecting the anaplasma phagocytophilum; the sequences SEQ ID NO:4 and SEQ ID NO:5 respectively refer to a forward primer and a reverse primer for detecting Ehrlichia chaffeensis; and the sequence SEQ ID NO:6 refers to a fluorescence probe for detecting the Ehrlichia chaffeensis. The invention also provides the kit for detecting the anaplasma phagocytophilum and Ehrlichia chaffeensis. The primer and probe provided by the invention are extremely high in specificity, two viruses can be detected at a time, and the efficiency is improved.

The invention discloses a PCR primer pair for rodent-borne disease pathogenic microorganisms. The primer pair is used for amplifying specific genes of leptospira interrogans, Rickettsi Mooseri, Orientia tsutsugamushi, anaplasma phagocytophilum, francisella tularensis, Coxiella burnetii and Bartonella. By adopting the PCR primer pair, on the basis of the TSP principle, the method for high-sensitiveand specific independent or combined detection of the seven kinds of pathogene.

Disclosed is the cloning, expression and purification of a hemolysin protein and its protein fragments in Anaplasma phagocytophilum. The recombinant hemolysin and its protein fragments are useful in the ELISA detection of anaplasma pathogen. The use of same as a kit for ELISA is also disclosed.

The invention discloses a PCR specific primer and molecular beacon TaqMan probe combination for simultaneously detecting two anaplasma and a detection method. The 5' end of a probe of anaplasma capraand anaplasma phagocytophilum is connected with a fluorescence reporting group (FAM or Cy5), the 3' end of the probe of the anaplasma capra and the anaplasma phagocytophilum is connected with a fluorescence quenching group (BHQ3 or BHQ1), the size of an amplification product of a special primer of the anaplasma capra groEL gene locus is 153 bp, and the size of an amplification product of a specialprimer of the anaplasma phagocytophilum 16S rRNA gene locus us 81 bp. A dual real-time quantitative PCR reaction system is 25 [mu]L of a PCR reaction system, and comprises 12.5 [mu]L of Premix Ex Taq, 0.4 [mu]M of each primer group, 0.4 [mu]M of probe DNA, 2 [mu]L of a template, and sterile deionized water is complemented to 25 [mu]L. The reaction condition of the dual real-time quantitative PCRamplification is a two-step method: 95 DEG C 30s, 95 DEG C 5s, 55.5 DEG C 30s, and operation is circulated for 40 times. The advantages of high detection speed, high sensitivity, high specificity, high efficiency, low cost and the like are achieved, and especially the absolute quantification of pathogens in a sample can be achieved.

The invention discloses a method for detecting anaplasma phagocytophilum employing a loop-mediated isothermal amplification (LAMP) technology. The detection method comprises an LAMP primer design, building of an LAMP reaction system and adoption of an LAMP detection method. Specificity detection, sensitivity detection and fluorescent visual detection are adopted in the LAMP detection method. The anaplasma phagocytophilum 16SrRNA is taken as a target gene locus, and has the characteristics of being fast, sensitive, efficient, low in cost and the like.

Axenic media and methods for growing E. Chaffeensis and / or A. phagocytophilum are provided. In general, the axenic media includes intracellular phosphate buffer (IPB), a carbon source, FBS, a mixture of amino acids, and at least one further component selected from the group consisting of glucose 6-phosphate (G6P), ATP, DTT, GTP, UTP, CTP, and any combination thereof.

The invention provides a preparation method of an anaplasma phagocytophilum histone antigen. The preparation method comprises the following steps: carrying out reverse transcription and PCR (Polymerase Chain Reaction) amplification on a gene segment containing a p44 gene, recovering and cloning an amplification product, extracting and reconfirming a target gene, and synthesizing a recombinant protein antigen rP44-47E. According to the preparation method of the anaphagophilic recombinant protein antigen, the recombinant protein antigen rP44-47E can be singly, efficiently and massively prepared; the prepared recombinant protein antigen rP44-47E is high in expression quantity and stability, the defects that p44 whole genome antigen is low in expression quantity and not easy to prepare are overcome, and the prepared recombinant protein antigen rP44-47E has the advantages of being high in specificity, capable of effectively reducing the detection cost and easy to standardize when serving as antigen protein.

The invention provides a recombinant antigen protein rP44-60 for detecting granulocytoplasmosis and a kit containing the antigen, and belongs to the technical field of immunology, and the recombinant antigen protein comprises an amino acid sequence as shown in SEQ ID NO.1 in a sequence table. The kit comprises a recombinant antigen protein rP44-60, a sample pad, a combination pad, an absorption pad, a nitrocellulose membrane, a colloidal gold probe, mouse IgG and goat anti-mouse IgG. The recombinant antigen protein disclosed by the invention is high in sensitivity and strong in specificity, and can be used for rapidly detecting an antibody in anaplasma phagocytophilum infected serum, so that the problem that the antibody is difficult to capture due to antigen variation of HGA is solved, the defect of existing antigen missing detection is made up, and the missed diagnosis rate is effectively reduced; according to the preparation method, cross reactivity caused by irrelevant sequences is reduced, and the expression quantity and stability of recombinant antigen protein are high; the kit has the characteristics of rapidness, sensitivity, high specificity, no limitation of experimental conditions and the like, and has low requirements on detection conditions.