30results about How to "No need for electrophoretic detection" patented technology

The invention provides LAMP (Loop-mediated Isothermal Amplification) primers for rapidly assaying tea tree anthracnose pathogens and an assay method. Aimed at the beta-tubulin(TUB2) gene sequence of tea tree anthracnose pathogens, the invention designs and screens out a set of specific assay primer group, and the primer group consists of four specific primers F3, B3, FIP and BIP. The primers screened out by the invention have high sensitivity and strong specificity, the established assay method ha the advantages of high accuracy, strong specificity, easiness in operation, short assay time, simple instruments and equipment and the like, and is applicable to the high-sensitivity rapid assay, identification and early disease diagnosis of tea tree anthracnose pathogens in diseased tissues andpathogen-carrying plants, and for the early monitoring of tea tree anthracnose in agricultural production, the invention is of great significance for the determination of optimal prevention and control time.

The invention discloses a loop isothermal amplification primer for quickly detecting tylenchulus semipenetrans and application of the loop isothermal amplification primer. The loop isothermal amplification primer comprises a primer pair TSF3 / TSB3 and a primer pair TSFIP / TSBIP, and the sequences of the primer pairs are shown as SEQ ID NO. 1 to 4 respectively. A loop isothermal amplification method is established through the loop isothermal amplification primer, loop isothermal amplification is performed through taking a sample DNA as a template, and results can be judged in two ways after the end of reaction, wherein the first way is that an amplification primer is subjected to electrophoresis, and a sample of a specific ladder-shaped strip, which occurs, is judged to be positive; the second way is that through adding SYBR green I into a reaction system, a sample of which the color is changed is observed with naked eyes to be positive. The loop isothermal amplification method is low in requirement for instruments and equipment, quick, safe, and high in specificity and sensitivity; a technical support is provided for the detection of the tylenchulus semipenetrans, in particular to the quick detection work of a grass-root quarantine unit for the tylenchulus semipenetrans, and the popularization and application values are very high.

The invention discloses a real-time recombinase-mediated isothermal amplification nucleic acid kit for rapid detection of toxoplasma gondii and an application thereof. The kit comprises standards. Thestandards are a positive plasmid containing a 529 gene sequence, specific primers and probes designed for 529 gene of the toxoplasma gondii. The primers designed based on a specific conservative target sequence of the 529 gene of the toxoplasma gondii for qualitatively detecting the 529 gene of the toxoplasma gondii in tissues, feces or blood samples comprise an upstream primer and a downstream primer. According to the present invention, a real-time fluorescent RAA technology is used to establish the method for rapid detection of the toxoplasma gondii. Compared with fluorescent quantitative PCR, the method has the advantages of low cost, simpleness and fastness, can conduct detection at constant temperature of 36 DEG C within 20min, and directly read detection results by a portable instrument. The method of the present invention achieves minimum detection size for toxoplasma gondii genomes up to 102 copies, has sensitivity comparable to the traditional fluorescent quantitative PCR, and is suitable for rapid diagnosis of clinical samples and laboratories.

The invention discloses an RAA constant temperature fluorescence detection method and a detection kit for yellow head virus (YHV) of shrimps. The detection kit comprises a forward primer SEQ ID NO.1,a reverse primer SEQ ID NO.2, a specific fluorescent probe SEQ ID NO.3, a reaction solution, reverse transcriptase, recombinant polymerase and a reference substance. The kit disclosed by the inventionhas the advantages of high specificity, high detection sensitivity as high as 0.10fg / mu L, high accuracy, reliability, simplicity and rapidness in operation, suitability for field detection and broadapplication prospect.

The invention discloses an RAA constant-temperature fluorescence detection method and detection kit for a grass carp reovirus type 2 (GCRV-2). The detection kit comprises a forward primer SEQ ID NO.1,a reverse primer SEQ ID NO.2, a specific fluorescent probe SEQ ID NO.3, a reaction solution, reverse transcriptase, recombinant polymerase and a reference substance. The kit disclosed by the invention is high in specificity; the detection sensitivity is high and can reach 1 fg / [mu]L; the accuracy is high and reliable; and the operation is simple and quick, suitability of field detection is achieved and the application prospect is wide.

The invention belongs to the technical field of molecular biology and discloses an RAA (recombinase aided amplification) constant temperature fluorescent detection primer probe set, kit and method forAfrican swine fever virus CD2V gene. A nucleotide sequence of a forward primer of the primer probe set is shown as SEQ ID NO. 1, a nucleotide sequence of a reverse primer is shown as SEQ ID NO. 2, and a nucleotide sequence of a specific fluorescent probe is shown as SEQ ID NO. 3, wherein a fluorescent reporter group is marked in the 5' end direction of the specific fluorescent probe, while a fluorescent quenching group is marked in the 3' end direction of the specific fluorescent probe. The primer probe set and kit provided by the invention help rapidly, conveniently, efficiently and specifically detect whether a sample is infected with an African swine fever wild strain or a CD2V deleted strain under an isothermal condition, do not need complex instruments and professional equipment on site, and are suitable for POCT (point-of-care testing).

The invention discloses recombinase-aid amplification (RAA) constant-temperature fluorescence detection method and reagent for Koi herpesvirus (KHV). The RAA constant-temperature fluorescence detection reagent for the KHV comprises a forward primer SEQ ID NO. 1, a reverse primer SEQ ID NO. 2, a specific fluorescent probe SEQ ID NO. 3, a reaction solution, recombinant polymerase and a reference substance. The RAA constant-temperature fluorescence detection reagent for the KHV disclosed by the invention is high in specificity, high in detection sensitivity (up to 100fg / microliter), high in accuracy and reliable; and moreover, the reagent is easy and convenient to operate as well as suitable for field testing. Thus, wide range of application scenarios is ensured.

The invention discloses a prawn covert mortality nodavirus (CMNV) RAA constant temperature fluorescence detection method and a prawn covert mortality nodavirus (CMNV) detection kit, wherein the detection kit comprises a forward primer SEQ ID NO.1, a reverse primer SEQ ID NO.2, a specific fluorescent probe SEQ ID NO.3, a reaction solution, reverse transcriptase, recombinant polymerase and a controlsubstance. According to the present invention, the kit has characteristics of strong specificity, high detection sensitivity, high accuracy, reliability, simple and rapid operation and wide application scene, and is suitable for on-site detection, wherein the detection sensitivity can reach 100 fg / [mu]L.

The invention discloses an isothermal amplification primer group capable of rapidly detecting cattle and sheep akabane disease viruses and application thereof. Nucleotide sequences of the isothermal amplification primer group are respectively shown as SEQIDNo.1-4. According to the invention, cDNA obtained through reverse transcription on RNA (ribonucleic acid) of a sample is taken as a template, the isothermal amplification primer is utilized for carrying out loop-mediated isothermal amplification detection, results can be judged in the following two ways after reaction is finished: electrophoresis is carried out on amplification products, and the sample with a specific stair-step strip is judged to be positive; calcein is added into a reaction system, judging is carried out by virtue of naked eyes, and the sample with change in colour is judged to carry positive cattle and sheep akabane disease viruses. The isothermal amplification method for detecting cattle and sheep akabane disease viruses by utilizing the isothermal amplification primer is low in requirement on instrument and equipment, rapid, safe, good in specificity and strong in sensitivity, provides a technical support for cattle and sheep akabane disease detection in the animal husbandry, especially rapid detection on cattle and sheep in a basic quarantine department, and has good popularization and application value.

The invention belongs to the technical field of molecular biology and discloses an RAA (recombinase aided amplification) constant temperature fluorescent detection primer probe set, kit and method forAfrican swine fever virus CD2V gene. A nucleotide sequence of a forward primer of the primer probe set is shown as SEQ ID NO. 1, a nucleotide sequence of a reverse primer is shown as SEQ ID NO. 2, and a nucleotide sequence of a specific fluorescent probe is shown as SEQ ID NO. 3, wherein a fluorescent reporter group is marked in the 5' end direction of the specific fluorescent probe, while a fluorescent quenching group is marked in the 3' end direction of the specific fluorescent probe. The primer probe set and kit provided by the invention help rapidly, conveniently, efficiently and specifically detect whether a sample is infected with an African swine fever wild strain or a CD2V deleted strain under an isothermal condition, do not need complex instruments and professional equipment on site, and are suitable for POCT (point-of-care testing).

The invention discloses a carp edema virus (CEV) RAA constant-temperature fluorescence detection method and a detection kit. The detection kit comprises a forward primer SEQ ID NO. 1, a reverse primerSEQ ID NO. 2, a specific fluorescent probe SEQ ID NO. 3, reaction liquid, recombinant polymerase and a reference substance. The kit of the invention has strong specificity; the detection sensitivityis high and can reach 2.02 fg / mu L; the accuracy is high and reliable; and the detection kit is simple, convenient and quick to operate, is suitable for field detection, and has wide application scenes.

The invention relates to a fluorescent quantitative PCR method for simultaneously detecting type 2 extracellular protein factor EF and hemolysin gene SLY of Streptococcus suis based on a SYBR Green dye melting curve analysis method. According to the conserved sequences of EF and SLY genes of Streptococcus suis type 2 published in GenBank, two pairs of specific primers were designed and synthesized, the PCR reaction system and reaction conditions were optimized, and the simultaneous detection of Streptococcus suis type 2 was established by analyzing the amplification curve and melting curve SYBR GreenI dye fluorescence quantitative PCR method for EF and SLY genes. On this basis, the molar ratio of EF and SLY primers was optimized, and the amplification efficiency of the two pairs of primers was adjusted, so that the peak sizes of the two products in the melting curve were close. The EF and SLY genes of Streptococcus suis type 2 can be detected conveniently and simultaneously by using the established SYBR Green dye melting curve analysis method.

The invention discloses a loop isothermal amplification primer for quickly detecting tylenchulus semipenetrans and application of the loop isothermal amplification primer. The loop isothermal amplification primer comprises a primer pair TSF3 / TSB3 and a primer pair TSFIP / TSBIP, and the sequences of the primer pairs are shown as SEQ ID NO. 1 to 4 respectively. A loop isothermal amplification method is established through the loop isothermal amplification primer, loop isothermal amplification is performed through taking a sample DNA as a template, and results can be judged in two ways after the end of reaction, wherein the first way is that an amplification primer is subjected to electrophoresis, and a sample of a specific ladder-shaped strip, which occurs, is judged to be positive; the second way is that through adding SYBR green I into a reaction system, a sample of which the color is changed is observed with naked eyes to be positive. The loop isothermal amplification method is low in requirement for instruments and equipment, quick, safe, and high in specificity and sensitivity; a technical support is provided for the detection of the tylenchulus semipenetrans, in particular to the quick detection work of a grass-root quarantine unit for the tylenchulus semipenetrans, and the popularization and application values are very high.

The invention discloses a ring isothermal amplification primer group capable of fast detecting pratylenchus neglectus. The primer group comprises an outer side primer pair pnF32 / pnB32 and an inner side primer pair of pnFIP2 / pnBIP2. The sequences of primers are shown as the SEQ ID NO.5-8. A ring isothermal amplification method is established according to the primer group, ring isothermal amplification is performed with sample DNA as a template, a result can be adjudged in two modes after reaction ends, according to one mode, electrophoresis is performed on amplification products, and samples with a specific ladder-like strip are considered as positive, and according to the other mode, SYBR green I is added into a reaction system, and samples where color changes are observed through naked eyes are considered as positive. The ring isothermal amplification method is low in requirement for instruments and equipment, fast, safe, good in specificity and high in sensitivity, the method provides technical support for detection of pratylenchus neglectus, particularly, a basic layer quarantine unit provides technical support for fast detection work of pratylenchus neglectus, and good application and popularization value is achieved.

The invention discloses a constant-temperature fluorescence detection method and a detection kit for Enterocystis hepatica (EHP) RAA of prawns. The detection kit includes a forward primer of SEQ ID NO.1, a reverse primer of SEQ ID NO.2, a specific fluorescent probe of SEQ ID NO.3, a reaction solution, a recombinant polymerase and a control substance. The kit of the invention has strong specificity; high detection sensitivity, which can reach 100 fg / μL; high accuracy and reliability; simple and quick operation, suitable for on-site detection, and has wide application scenarios.

The invention provides a LAMP primer and a detection method for rapidly detecting tea tree anthracnose bacteria. The present invention is aimed at tea tree anthracnose fungus beta-tubulin (TUB2) gene sequence designed and screened a set of specific detection primers, which consisted of 4 specific primers F3, B3, FIP and BIP. The primers screened by the present invention have high sensitivity and strong specificity, and the established detection method has the advantages of high accuracy, strong specificity, easy operation, short detection time, simple equipment, etc., and is suitable for tea tree anthracnose bacteria in diseased tissues and infected plants The high-sensitivity rapid detection, identification and early diagnosis of diseases are of great significance for the early monitoring of tea tree anthracnose disease in agricultural production and the determination of the best period of disease control.

The invention discloses a constant-temperature fluorescence detection method and a detection kit for shrimp infectious subcutaneous and hematopoietic necrosis virus (IHHNV) RAA. The detection kit includes a forward primer of SEQ ID NO.1, a reverse primer of SEQ ID NO.2, a specific fluorescent probe of SEQ ID NO.3, a reaction solution, a recombinant polymerase and a control substance. The kit of the invention has strong specificity; high detection sensitivity, which can reach 0.32fg / μL; high accuracy and reliability; simple and fast operation, suitable for on-site detection, and has wide application scenarios.

The invention discloses a loop-mediated isothermal amplification primer set for identifying three kinds of pratylenchus pratensis on sugarcane and a kit comprising the loop-mediated isothermal amplification primer set. The loop-mediated isothermal amplification primer set comprises three groups of primers, namely, an outer primer pair PZF3 / PZB3 and an inner primer pair PZFIP / PZBIP, an outer primer pair PPF3 / PPB3 and an inner primer pair PPFIP / PPBIP, as well as an outer primer pair PBF3 / PBB3 and an inner primer pair PBFIP / PBBIP, wherein the nucleotide sequences of the primers are respectively shown as SEQ ID NO.1-12. The three kinds of pratylenchus pratensis are pratylenchus zeae, P. parazeae and P. brachyurus. The loop-mediated isothermal amplification primer set and the kit comprising the loop-mediated isothermal amplification primer set have low requirements on instrument and equipment, are simple, rapid and safe, and are good in specificity and high in sensitivity, and a technical support is provided for the detection for pratylenchus zeae, P. parazeae and P. brachyurus, especially, the rapid identification and distinguishing for the three kinds of pratylenchus pratensis of the grassroots units.

The invention discloses a shrimp iridescent virus (SIV) RAA constant temperature fluorescence detection method and a detection kit. The detection kit includes a forward primer of SEQ ID NO.1, a reverse primer of SEQ ID NO.2, a specific fluorescent probe of SEQ ID NO.3, a reaction solution, a recombinant polymerase and a control substance. The kit of the invention has strong specificity; high detection sensitivity, which can reach 2fg / μL; high accuracy and reliability; simple and fast operation, suitable for on-site detection, and has wide application scenarios.

The invention discloses an infectious hematopoietic necrosis virus (INHV) RAA (recombinase-aid amplification)thermostaticfluorescence detection method and a detection kit. The detection kit includes aforward primer SEQ ID NO. 1,a reverse primer SEQ ID NO. 2,a specific fluorescent probe SEQ ID NO. 3,reaction liquid, reverse transcriptase, recombinant polymerase, and reference substance. The detection kit is high in specificity; detection sensitivity is high and can reach 0.1fg / [mu]L; high accuracyand reliability are realized; operation is easy, convenient, and quick, and the detection kit is suitable for field testing and has a wide range of application scenarios.

The invention discloses a group of isothermal amplification primers for rapid detection of cattle and sheep Akabane disease virus and application thereof. The nucleotide sequences of the primers are respectively as SEQ? ID? Shown in NO.1~4. In the present invention, the cDNA obtained by sample RNA reverse transcription is used as a template, and the above-mentioned primers are used for loop-mediated isothermal amplification detection. After the reaction, the result can be judged in two ways: one is that the amplified product is subjected to electrophoresis, and a specific ladder-like appearance occurs. The sample of the strip is judged as positive; the second is by adding calcein to the reaction system, and judged by naked eyes, the sample with color change is judged as positive for bovine and sheep Akabane disease virus. The isothermal amplification method for detecting Akabane disease virus in cattle and sheep by using the primers in the present invention has low requirements on instruments and equipment, is fast, safe, has good specificity, and has strong sensitivity. The quarantine unit has provided technical support for the rapid detection of cattle and sheep, which has very good promotion and application value.