51results about How to "Meet on-site inspection" patented technology

The invention belongs to the field of a genetic engineering, and relates to a loop-mediated isothermal amplification (LAMP) primer composition for detecting phytophthora rot and an application thereof. The primer composition comprises a forward inner primer FIP showed in SEQ ID NO.1, a backward inner primer BIP showed in SEQ ID NO.2, a forward outer primer F3 showed in SEQ ID NO.3, a backward outer primer B3 showed in SEQ ID NO.4, and a reverse annular primer LB showed in SEQ ID NO.5. The LAMP primer composition can be applied to detection or identification of phytophthora rot. The LAMP primer composition has the advantages of good sensibility for specificity, rapid amplification, high efficiency and simple identification in the application of detecting phytophthora rot.

The invention discloses a method and a primer composition for detecting soybean fusarium oxysporum based on an LAMP (loop-mediated isothermal amplification) technology. The primer composition comprises four specific primers FIP, BIP, F3 and B3 for detecting the LAMP molecules of the soybean fusarium oxysporum, and two ring primers LF and LB used for increasing the reaction speed. The method is used for performing LAMP on DNAs to be detected by the primer composition provided by the invention; a detection result is observed by naked eyes or under the irradiation of ultraviolet light with the wavelength of 245 nm; the color change and the fluorescence intensity of SYBR Green I are taken as result judgment standards. The invention provides the new molecular detection method and the primer composition for detecting the soybean fusarium oxysporum, the soybean fusarium oxysporum is subjected to LAMP detection, the detection cycle is short, the accuracy is high, the sensitivity is high, and the detection result is observed by the naked eyes.

The invention provides a LAMP detection primer set of broccoli stem rot fungus and a rapid detection method thereof. The LAMP primer set was composed of one pair of outer primers F3 / B3 and one pair ofinner primers FIP / BIP. The sequences of F3 / B3 and FIP / BIP primers were as follows: SEQ ID 1-4. Comprises the following steps of: 1) extracting DNA of a sample to be tested; 2) LAMP amplification; 3)detect that amplification product by using a fluorescent dye SYBR Green I for color development or agarose gel electrophoresis of the amplification result. So that that invention can quickly, Convenient, high efficiency, high specificity, high sensitivity detection of broccoli stem rot pathogen, does not require complex and expensive instruments, can better meet the rapid detection of broccoli stem rot pathogen in the field, can be widely used in grass roots units of early disease diagnosis and disease pathogen monitoring, identification, for the control of broccoli stem rot disease to providereliable technical and theoretical basis.

The invention discloses an LAMP detection primer composition for phytophthora infestans and an LAMP detection kit and an LAMP detection method of the LAMP detection primer composition. The LAMP detection primer composition consists of a positive inner primer FIP, a reverse inner primer BIP, a positive outer primer F3, a reverse outer primer B3 and a reverse ring primer LB. The detection method disclosed by the invention is high in accuracy, high in specificity, convenient to operate and high in practicability, realizes constant-temperature multiplication, also provides a new technical platform for detection on the phytophthora infestans and can be used for performing high-sensitivity quick detection on the phytophthora infestans; meanwhile, pathogens can be identified at the initial stage of disease invasion, and the pathogens in field soil can be detected. The LAMP detection primer composition has an important significance for preventing and treating late blight of potatoes and tomatoes, reducing the blind use of pesticide, lowering the production cost and alleviating the environmental pollution of the pesticide.

The invention discloses botryosphaeria dothidea LAMP (Loop-mediated Isothermal Amplification) detection primers and application thereof. A primer composition for detecting botryosphaeria dothidea based on an LAMP technology consists of 6 specific primers, namely F3, B3, FIP, BIP, LF and LB. The invention further provides a botryosphaeria dothidea LAMP detection method. The botryosphaeria dothideaLAMP detection method comprises the following steps: 1), extracting DNA of a sample to be detected; 2), performing loop-mediated isothermal amplification on the DNA of the sample to be detected by using the primer composition provided by the invention; 3), performing result detection, wherein when a chromogenic result is green fluorescence or a trapezoidal electrophoretic band appears, a positiveresult is determined, and when a chromogenic result is orange or orange yellow or no trapezoidal electrophoretic band appears, a negative result is determined. A novel molecular detection method and the novel primer composition are provided for the botryosphaeria dothidea, the botryosphaeria dothidea can be detected quickly, conveniently and efficiently at high specificity and high sensitivity, and a reliable technological and theoretical basis is provided for early diagnosis, detection and rapid import and export inspection of the botryosphaeria dothidea.

The invention provides a primer combination for detecting succulent plant stem fusarium moniliforme based on LAMP (Loop-mediated Isothermal Amplification) and application thereof. The LAMP primer combination for detecting the succulent plant stem fusarium moniliforme consists of 1 pair of outside primers F3 / B3 and a pair of inside primer FIP / BIP, wherein the F3 / B3 and FIP / BIP primer sequences areshown as SEQ ID NO. 1 to 4. By adopting the detection system disclosed by the invention, the succulent plant stem fusarium moniliforme can be detected rapidly, conveniently and efficiently at high specificity and high sensitively through incubation at the constant temperature of 65 DEG C for 1 hour without complex and expensive instruments, and in-situ detection of the succulent plant stem fusarium moniliforme can be met well. The detection system can be widely applied to early diagnosis of field diseases of establishment units as well as monitoring and identification of viruses, and a reliable technological and theoretical basis is provided for prevention and control of the succulent plant stem fusarium moniliforme.

The invention discloses an LAMP primer composition, kit and detection method for detecting pythium ultimum. The primer composition comprises a positive inner primer FIP, a reverse inner primer BIP, a positive outer primer F3, a reverse outer primer B3 and a positive loop primer LP. The genome DNA of a to-be-detected microorganism serves as a template, and the LAMP primer composition serves as a primer for LAMP reaction; then the color change of an amplification product is observed, if the color changes from purple to sky blue, it represents that pythium ultimum exists in the to-be-detected microorganism, and if the color does not change and is still purple, it represents that pythium ultimum does not exist in the to-be-detected microorganism. Compared with a traditional detection technology that pythium ultimum is verified according to morphological characteristics, the method has higher accuracy, sensitivity and effectiveness, is convenient to operate and good in practicality, provides a new technical platform for pythium ultimum detection, and can be used for high-sensitivity quick detection of pythium ultimum.

The invention discloses a primer composition for detecting pythium arrhenomane based on a loop-mediated isothermal amplification technique and a detection method thereof. The detection method comprises the following steps of extracting a genomic DNA (Deoxyribonucleic Acid) of a to-be-detected microbe, taking a genomic DNA solution as a reaction template, and adding a detection solution in an LAMP(Loop-Mediated Isothermal Amplification) kit to carry out an LAMP reaction, wherein the LAMP reaction comprises the procedures of reacting at 64 DEG C for amplification for 60min, afterwards, observing the change of the color of an amplification product, if the color becomes from purple to sky blue, indicating that the pythium arrhenomane exists in a to-be-detected matter, and if the color is notchanged or is still purple, indicating that the pythium arrhenomane does not exist in the to-be-detected matter. Compared with a conventional detection technique of authenticating the pythium arrhenomane according to a morphological character, the detection method has higher accuracy, sensitivity and effectiveness, moreover, is convenient to operate and good in practicability, is used for providing a new technical platform for the detection of the pythium arrhenomane, and can be used for the quick detection of the pythium arrhenomane.

The invention discloses a nucleic acid (DNA / RNA) real-time constant-temperature gene amplification detection method. The method is a rapid and efficient novel isothermal amplification detection method taking microplate reader detection as a core. The method particularly comprises the following steps that a to-be-detected sample DNA / RNR template is added into a 15 microliters-25 microliters of LAMP amplification system with certain neutral red concentration and then put into a microplate reader with the temperature ranging from 60 DEG C to 65 DEG C to detect change of absorbance of a reaction system in real time, and therefore the reaction is indirectly monitored. A reaction signal is expressed in the form of a curve, the curve enters a logarithmic phase within 30 min-60 min to express that a detected sample is positive, and the curve extends all the time but not enters the logarithmic phase to express that the detected sample is negative. The nucleic acid (DNA / RNA) real-time constant-temperature gene amplification detection method is simple, rapid and convenient, a great push function is achieved for subsequent basic-level popularization, compared with a traditional PCR detection method, rapid and safe detection of a gene is achieved, and an important significance in the aspects of blocking transmission routes, effectively controlling and tracking infection sources and effectively controlling outbreak and prevalence of superbacteria is achieved.

The invention discloses a carbendazim resistant genotype F200Y sclerotinia sclerotiorum LAMP detection primer composition and a LAMP test kit and a LAMP test method. The LAMP detection primer composition comprises a forward inner primer FIP, a backward inner primer BIP, a forward outer primer F3 and a backward outer primer B3. The test method is simple and easy, good in practicability, high in sensitivity, strong in specificity, and high in accuracy, and can achieve the isothermal amplification, a new technology platform is provided for testing carbendazim resistant genotype F200Y sclerotinia sclerotiorum, and the method can be used for rapid high sensitivity testing of the carbendazim resistant genotype F200Y sclerotinia sclerotiorum, monitoring of sclerotinia sclerotiorum carbendazim resistant group, and timely understanding of the development trend of the resistance group, and the detection primer composition has great significance to crop sclerotiniose resistance management and scientific guiding drug use, production cost reduction and deduction of pesticide pollution of the environment.

The invention discloses an LAMP detection primer composition for phytophthotacactorum, as well as an LAMP detection kit and an LAMP detection method of the LAMP detection primer composition. The LAMP detection primer composition consists of a positive inner primer FIP, a reverse inner primer BIP, a positive outer primer F3, a reverse outer primer B3 and a reverse loop primer LB; detailed sequences of all the primers are as follows: FIP: 5'-TCTGGGCACAACCGCAAAAA-TTTGCGAGCTCCAGATTTCC-3'; BIP: 5'-AATCCGTACGATCGAGCTGGAC-ACACGCCACGTCTGCT-3'; F3: 5'-TTCTGCGCTAGGCGACC-3'; B3: 5'-CACACAAGTGGACCGTTAG3'; LB: 5'-GGAAAGACCATCAAGCTCCAGAT-3'. The detection method disclosed by the invention is high in accuracy, high in specificity, convenient to operate and good in practicability; constant-temperature amplification is realized; meanwhile, a novel technology platform is provided for detection of the phytophthotacactorum; the detection method can be applied to high-sensitivity rapid detection of the phytophthotacactorum; simultaneously, pathogens can be identified in initial stage of invasion of diseases, and the phytophthotacactorum soil of a field can be detected. The LAMP detection primer composition for the phytophthotacactorum, as well as the LAMP detection kit and the LAMP detection method of the LAMP detection primer composition, disclosed by the invention, play an important role in epidemic treatment caused by the phytophthotacactorum, reduction of sightless usage of pesticides, reduction of production cost and reduction of environmental pollution caused by the pesticides as well.

The invention discloses a method for molecular detection of Phytophthora cambivora(Petri)Buisman with the loop-mediated isothermal amplification (LAMP) technique based on color differentiation, and primers. The sequences of the primers are shown from SEQ ID NO.1 to SEQ ID NO.6. According to the detection system, under the isothermal condition of 62 DEG C, Phytophthora cambivora(Petri)Buisman can be detected quickly, conveniently and efficiently with high specificity and sensitivity, no complicated instrument is needed, a new technical platform is provided for detection of Phytophthora cambivora(Petri)Buisman, and the requirement for field detection of Phytophthora cambivora(Petri)Buisman can be well met. The detection system is suitable for inspection and quarantine of entry / exit plants and plant products, and investigation, rapid diagnosis and monitoring of diseases, and is of great importance in preventing Phytophthora cambivora(Petri)Buisman from entering China. Meanwhile, the establishment of the system provides technical guidance and theoretical basis for detection of other pathogenic bacteria.

The invention discloses Phytopythiumvexans detection primers, LAMP detection system, kit and method. The detection primers include a forward inner primer FIP, a backward inner primer BIP, a forward outer primer F3, a backward outer primer B3 and a forward loop primer LF. Compared with a traditional detection technology for identifying Phytopythiumvexans according to morphological characteristics,the detection method provided by the invention has the advantages of high accuracy and sensitivity and convenience in operation, and provides a new technical platform for Phytopythiumvexans detectionwhile realizing constant-temperature amplification. The method can be used for high-sensitivity rapid detection of the Phytopythiumvexans in production practice, pathogens can be identified in the early stage of disease infection, and the Phytopythiumvexans in field soil can be detected. According to the invention, reliable technical and theoretical bases are provided for prevention and treatmentof the Phytopythiumvexans.

The invention discloses a primer combination for detecting Phialophora gregata by loop-mediated isotherm amplification technique and application thereof. The primer combination is composed of a forward outer primer F3 shown in SEQ ID NO.1, a reverse outer primer B3 shown in SEQ ID NO.2, a forward inner primer FIP shown in SEQ ID NO.3, a reverse inner primer BIP shown in SEQ ID NO.4, a loop primer LF shown in SEQ ID NO.5, and a loop primer LB shown in SEQ ID NO.6. The primer combination can be applied to preparation of a Phialophora gregata loop-mediated isotherm amplification detection kit. The kit has good specificity and sensitivity, has high amplification speed, has high efficiency, can identify conveniently, can meet the present crying need for rapid detection of the Phialophora gregata, can be used for quarantine of imports and exports, field quarantine and other field testing, and can be easily promoted and applied in a large range.

The invention discloses a LAMP detection primer combination of eugenopsis and a LAMP detecting kit and method thereof. The loop-mediated isothermal amplification detection primer combination is composed of a forward inner primer FIP, a backward inner primer BIP, a forward outer primer F3 and a backward outer primer B3. The detecting method is high in accuracy and specificity, convenient to operateand high in practicability, constant-temperature amplification is realized, a new technological platform is provided for detection of quarantine eugenopsis, the method can be used for high-sensitivity rapid detection of eugenopsis, pathogens are authenticated in the early stage of disease infection, and eugenopsis in import and export fruit samples can be detected. The method has important significance in reducing diseases caused by eugenopsis, reducing the blind use of pesticides, reducing production cost and reducing pesticide environment pollution.

The invention discloses a method for detecting gene F200Y of fusarium graminearum resisting carbendazim on basis of a loop-mediated isothermal amplification technology and a primer composition. The primer composition for detecting carbendazim-resisting gene-type F200Y fusarium graminearum consists of a forward inner primer FIP shown as SEQ ID NO.2, an inverse inner primer BIP shown as SEQ ID NO.3, a forward outer primer F3 shown as SEQ ID NO.4 and a forward outer primer B3 shown as SEQ ID NO.5. The detection method is simple, easy, good in practicability, high in sensitivity, high in specificity, high in accuracy and capable of realizing isothermal amplification, a novel technical platform is provided for the detection of the carbendazim-resisting gene-type F200Y fusarium graminearum, the carbendazim-resisting colony of fusarium graminearum can be monitored, the development state of the resistance colony can be known in time, and the method and the primer composition have important significance for governing the drug resistance of wheat scab and scientifically instructing the drug application, reducing the production cost and reducing the environmental pollution of chemicals.

The invention discloses a method for fast detecting botryis cinerea's resistance to QoI type bactericides and a primer composition. The method is based on the loop-mediated isothermal amplification technology. The LAMP detecting primer composition comprises a forward inner primer FIP, a backward inner primer BIP, a forward outer primer F3 and a backward outer primer B3. The method has the advantages that the method is simple and convenient, good in practicality, high in sensitivity, high in specificity and high in accuracy, isothermal amplification is realized, a new technical platform is provided to the detection of the botryis cinerea's resistance to the QoI type bactericides, the QoI type bactericide resistance groups of botryis cinerea can be monitored, and the development trend of the resistance groups can be learned about timely; the method has important practical significance to manage crop grey mold resistance, reduce pesticide environmental pollution and lower agricultural production cost.

The invention discloses a primer pair and a probe for detecting a swine-derived component, and discloses a kit designed according to the primer pair and the probe. The kit comprises an RAA reaction system and an LFD test strip; the RAA reaction system comprises RAA reaction general dry powder, a Tris-HCl buffer solution, an upstream primer ZHU-F, a downstream primer ZHU-R, a probe ZHU-PRO, a sample DNA extracting solution, MgAcO and ddH2O; and an avidin-colloidal gold specific antibody is arranged on a quality control line of the LFD test strip, and a biotin antibody and a fluorophore antibody are arranged on a detection line of the LFD test strip. On the basis of a recombinase-mediated isothermal nucleic acid amplification technology and a lateral flow immunoassay technology, a novel swine-derived component adulteration field detection method is provided, visual detection of naked eye results can be achieved, compared with other detection methods, the method is high in detection sensitivity, easy, convenient and rapid to operate, and does not need special equipment, the results are accurate, the detection of field samples can be met, and the method is especially suitable for field detection in farmer markets, supermarkets, wholesale markets and the like.

The invention discloses a recombinant plasmid. The recombinant plasmid is obtained by inserting a sef14 operon gene into an expression vector pBR322. The invention also discloses a recombinant strain.The recombinant strain is obtained by introducing the recombinant plasmid into inert carrier bacteria S9. The invention also discloses a preparation method and an application of the recombinant strain. According to the invention, SEF14 is subjected to surface display on the inert carrier bacterium S9 for the first time to express single fimbriae SEF14, so that background non-specific reaction canbe avoided, salmonella enteritidis infection can be specifically detected at the same time, the method has the advantages of rapidness, specificity, sensitivity, simplicity, low cost and the like, and the requirements of on-site and large-scale detection can be met. The SEF14 functional bacterial hair can be used for detecting and monitoring salmonella enteritidis infected chicken flocks after single bacterial hair SEF14 is shown and expressed on the upper surface of an inert carrier, only 5-10 microliters of the detection sample and an isopyknic detection reagent are needed, the salmonella enteritidis infected chicken flocks can be detected and monitored by utilizing a simple glass plate agglutination reaction, and observing of a reaction result by naked eyes on site and accurately judging whether an animal is infected by salmonella enteritidis or not are carried out.

The invention discloses a primer pair and a probe for detecting a sheep-derived component, and discloses a kit designed according to the primer pair and the probe. The kit comprises an RAA reaction system and an LFD test strip; the RAA reaction system comprises RAA reaction general dry powder, Tris-HCl buffer solution, an upstream primer YANG-F, a downstream primer YANG-R, a probe YANG-PRO, sample DNA extracting solution, MgAcO and ddH2O; and an avidin-colloidal gold specific antibody is arranged on a quality control line of the LFD test strip, and a biotin antibody and a fluorophore antibody are arranged on a detection line of the LFD test strip. According to the primer pair and the probe for detecting the sheep-derived component, a novel sheep-derived component adulteration field detection method is provided based on a recombinase-mediated isothermal nucleic acid amplification technology and a lateral flow immune technology, so that visual detection of naked eye results can be realized, and compared with other detection methods, the method is high in detection sensitivity, simple, convenient and rapid to operate, does not need special equipment, and the method is especially suitable for field detection in farmer markets, supermarkets, wholesale markets and the like.

The present invention discloses a novel target Phibe_s00003g00002.1 for specifically detecting phytophthora hibernalis, and a specific detection primer of the novel target and a detection method. Thedetection target Phibe_s00003g00002.1 has a DNA sequence as shown in SEQ ID NO: 1, and a protein sequence as shown in SEQ ID NO: 2. Meanwhile, the invention discloses a combination of a specific primer and a probe for RPA detection technology based on the specific detection target Phibe_s00003g00002.1. A forward primer sequence of the specific primer is shown in SEQ ID NO: 3, a reverse primer sequence of the specific primer is shown in SEQ ID NO: 4, and a probe sequence is shown in SEQ ID NO: 5. According to the invention, the novel detection target is discovered. The rapid detection method based on the target has high accuracy, high specificity, convenient operation and good practicability, and achieves constant temperature amplification. Meanwhile, the combination of the primer and the probe designed by the invention is used for RPA-LFD detection technology, has high specificity and high sensitivity, and provides a new technical platform for the detection of phytophthora hibernalis.

The invention discloses an LAMP (Loop-mediated Isothermal Amplification) primer combination for detecting tomato phytophthora infestans and application. The LAMP primer combination for detecting the tomato phytophthora infestans consists of four specific primers, and the primer combination disclosed by the invention is applied in the LAMP detection of the tomato phytophthora infestans. A detection system disclosed by the invention is incubated under the isothermal condition of 64 DEG C for 1 hour, SYBR green I developer is adopted for detection by development, and with whether green fluorescence is observed as a standard, a detection result is judged. The LAMP primer combination has the advantages of high accuracy, high specificity, convenience in operation and high practicability, does not need complex instruments, realizes isothermal amplification, provides a novel molecular detection technique for the tomato phytophthora infestans, can be used in rapid, sensitive and accurate detection in the early stage or infection incubation of the tomato phytophthora infestans in production practice, and provides a reliable technical and theoretical basis for the prevention and treatment of the tomato phytophthora infestans.

The invention discloses a primer pair and probe for detecting chicken-derived components, and discloses a kit designed according to the primer pair and probe. The kit comprises an RAA reaction system and an LFD test strip, wherein the RAA reaction system comprises general dry powder for an RAA reaction, a Tris-HCl buffer solution, an upstream primer JI-F, a downstream primer JI-R, a probe JI-PRO, a sample DNA extracting solution, MgAcO and ddH2O; a quality control line of the LFD test strip is provided with an avidin-colloidal gold specific antibody; and a detection line of the LFD test strip is provided with a biotin antibody and a fluorophore antibody. Based on a recombinase-mediated isothermal nucleic acid amplification technology and a lateral flow immunoassay technology, the invention provides a novel chicken-derived component adulteration on-site detection method; visual detection of naked eye results can be realized; and compared with other detection methods, the method is high in detection sensitivity, simple and rapid to operate, free of special equipment and accurate in result, can meet the detection requirement of field samples, and is especially suitable for field detection of farmer markets, supermarkets, wholesale markets and the like.

The invention discloses a new detection target of Phytophthora syringae, Psyri_s00018g00015.1, a detection primer, a detection kit and a detection method. The protein sequence of the detection target Psyri_s00018g00015.1 is shown in SEQ ID NO: 1, The DNA sequence encoding this protein is shown in SEQ ID NO:2. At the same time, the present invention also discloses the specific primer and probe combination of the RPA detection technology for the specific detection target Psyri_s00018g00015.1, the forward primer sequence is as shown in SEQ ID NO: 3, and the reverse primer sequence is as shown in SEQ ID NO: 4, the probe sequence is shown in SEQ ID NO:5. The invention discovers a new detection target of Phytophthora syringae, provides a new detection method for the detection of Phytophthora syringae, and at the same time, the RPA-LFD detection primers and probes developed based on the target can realize the specific detection of Phytophthora syringae, high sensitivity.

The invention discloses an LAMP (Loop-Mediated Isothermal Amplification) combined detection primer, a detection kit and a detection method for rhizome traditional Chinese medicinal materials. The LAMP combined detection primer comprises a forward outer primer F3, a reverse outer primer B3, a forward inner primer FIP, a reverse inner primer BIP, a forward loop primer LF and a reverse loop primer LB; the detection kit comprises an LAMP detection primer and an LAMP reaction solution. The detection method has high accuracy and sensitivity and is convenient to operate, isothermal amplification is achieved, and meanwhile a new technical platform is provided for detection of common root rot and southern blight pathogenic bacteria. The kit can be used for high-sensitivity rapid detection of common root rot and southern blight pathogenic bacteria of traditional Chinese medicinal materials in production practice, can detect pathogenic bacteria in the early stage of disease infected plants, and can detect pathogenic bacteria in field soil. Reliable technical and theoretical basis is provided for prevention and treatment of common root rot and southern blight of rhizome traditional Chinese medicinal materials (such as salvia miltiorrhiza, radix aconiti carmichaeli and rhizoma corydalis).