The invention discloses a real-time isothermal amplification-based kit for human papilloma virus E6/E7 gene detection, and application thereof. A primer and a probe are designed by aiming at 14 high-risk HPV, and rapid detection and identification on HPV E6/E7 mRNA can be realized specifically. The kit provided by the invention has the characteristics of high speed, high efficiency, sensitivity, specificity, real-time detection and analysis and the like, provides a rapid and accurate molecular detection method for generation investigation of HPV and prevention and treatment of cervical cancer, greatly reduces the inspection cost and has an important popularization and application value.

Real-time isothermal amplification-based kit for human papilloma virus E6/E7 gene detection, and application thereof

The invention discloses a primer combination for LAMP (loop-mediated isothermal amplification) detection and a reaction system thereof. The primer combination comprises a primer group for LAMP detection of HPV16 and a primer group for subtype-18 HPV (human papilloma virus) detection. The primer combination and the reaction system thereof disclosed by the invention have the following advantages andeffects that during a LAMP reaction in detection, a judgment result can be observed by naked eyes when green fluorescence appears in the reaction solution, and thus is more visual and clearer; the detection adopting the gene diagnosis method in the invention only takes 30 minutes; compared with the PCR (polymerase chain reaction) technology, the methodological indexes such as sensitivity, specificity and detection range are equivalent or superior to those of the PCR technology; field high-throughput rapid detection can be realized without depending on any special instrument; moreover, the detection cost is much lower than that of the fluorescence quantitative PCR technology, and the effects of short detection time, visual and clear judgment result, high sensitivity and strong specificityare realized.

Primer combination for LAMP (loop-mediated isothermal amplification) detection and reaction system thereof

The invention discloses a detection primer set for reverse transcription loop-mediated isothermal amplification (RT-LAMP) of Noroviruses (NV), a detection method and a kit. The detection primer set comprises an upstream outer primer F3, a downstream outer primer B, an upstream inner primer FIP and a downstream inner primer BIP, all of which are shown as the SEQ ID NO. 1, 2, 3 and 4 respectively. The RT-LAMP technology for detecting NV is simple, specific, sensitive and suitable for field detection and simple laboratory detection, and a novel virus detection method is provided. During sample detection, the sensitivity of the RT-LAMP technology reaches 97.7% higher than 90.7% of RT-PCR, and the RT-LAMP technology is more suitable for rapid detection of field samples and rapid detection in a simple laboratory and can be widely applied to hospitals, the food industry, entry and exit inspection and quarantine, quality supervision and other sectors and fields.

Detection primer set for reverse transcription loop-mediated isothermal amplification of Noroviruses, detection method and kit

The invention relates to a novel isothermal nucleic acid amplification technology, namely an isothermal multiple self-pairing triggering amplification technology (IMSA) According to the technology, amplification of a pathogeny gene can be realized under the condition of isothermality and under the action of a single functional enzyme without a thermal cycle instrument The result determination of the isothermal multiple self-pairing triggering amplification technology (IMSA) can be realized by simple measures such as observing white precipitate of by-product magnesium pyrophosphate, adding a chromogenic agent and observing the color change before or after amplification The most unique characteristic of the technology is that multiple oligonucleotide structures which can perform self-pairing and then trigger circular amplification will be generated during the process of amplification so that the triggering probability of following circular amplification is markedly increased, the amplification efficiency and detection sensitivity are promoted On the basis of the isothermal multiple self-pairing triggering amplification technology (IMSA) we establish detection method of hand-foot-and-mouth disease pathogen coxsackievirus type A16 (CVA16) and human enterovirus type EV71 (EV71) VP1 gene, and rapid detection method of human influenza a virus H7N9 HA gene By the application of the invention, simple, rapid, highly sensitive and highly specific detection can be realized

Novel isothermal multiple self-pairing triggering amplification technology

The invention discloses a constant-temperature fluorescence detection primer group, a kit and a detection method for avoiding false-negative hand-foot-mouth virus CoxA16. The primer group comprises a detection primer group and an internal standard primer group, wherein the detection primer group is designed according to a hand-foot-mouth virus CoxA16 CAA gene which has a sequence as shown in SEQ ID NO.1-6; and the internal standard primer group is designed according to a hand-foot-mouth virus CoxA16 CAB gene which as a sequence as shown in SEQ ID NO.7-12. Besides the primer groups, the constant-temperature detection kit for hand-foot-mouth virus CoxA16 further comprises DNA polymerase, a reaction liquid, reverse transcriptase, an internal standard target segment, positive control and negative control. The detection primer group, the kit and the detection method have the advantages of quickness, high efficiency, convenience in operation, high specificity, high sensitivity, field detection availability and the like, can be used for effectively avoiding occurrence of false negative, and is suitable for popularization and application.

Constant-temperature fluorescence detection primer group, kit and detection method for avoiding false-negative hand-foot-mouth virus CoxA16

The invention belongs to the biotechnological application field, and relates to the detection and the monitoring of Chinese-common six HPVs of cervical scraping smear specimens of suspected HPV (human papilloma virus) infected patients with genital damage (comprising genital warts and cervical epithelial neoplasia) as main symptoms in disease prevention control mechanisms at all levels, papilloma virus detection network laboratories, sentinel point hospitals and the like. The conserved sequences of E6, E7 or L1 genes of six HPV subtypes are analyzed by bioinformatics professional software through applying gene color based loop-mediated isothermal amplification (LAMP) technology, six primers of each subtype, which completely couple with eight binding domains in each target sequence, are designed, single reverse transcription and nested PCR secondary application steps are omitted, an HNB (hydroxynaphthol blue) dye is added before the reaction, the amplification in a 63DEG C isothermal environment for 1h can be completed only through one step, and a detection result can be determined through the color change observed with naked eyes and the white precipitate at the bottom of a tube. Compared with traditional PCR methods, the technology provided in the invention has the advantages of safety, specificity, sensitivity, and convenience.

Color based loop-mediated isothermal amplification technology for detecting HPV6, HPV11, HPV16, HPV18, HPV52 and HPV58

The invention relates to a method for detecting CA16 virus gene by color-based reverse transcriptase loop-mediated isothermal amplification, which belongs to the filed of biotechnology application and relates to a method for detecting and monitoring the human enterovirus CA16 type VP1 gene of such specimens as throat swabs, serum and excrement of fever patients with suspected hand-foot-mouth diseases which takes rash or herpes on hand, foot and oral cavity as main symptoms in all-level disease prevention and control institutions, enterovirus detection internet laboratories, sentinel hospitals and the like The advantages of the invention are specifically embodied as follows: analyzing the conserved sequence of the human enterovirus CA16 type VP1 gene with bioinformatics professional software by applying gene color based reverse transcriptase loop-mediated isothermal amplification (RT-LAMP), designing six primers which are completely matched with eight binding domains in the recognized VP1 target sequence, omitting the steps of single reverse transcription and secondary amplification of nested PCR, adding HNB (hydroxynaphthol blue) dyes before reaction, being capable of completing amplification for 1h in the isothermal environment at 65 DEG C only by one step and finally observing the color variation and white precipitates at the tube bottom with naked eyes to determine the detection result Compared with the traditional RT-PCR method, the method of the invention has the advantages of safety, specificity, sensitivity and convenience

Method for detecting CA16 virus gene by color-based reverse transcriptase loop-mediated isothermal amplification

Belonging to the field of biotechnology application, the invention relates to detection and monitoring of Norovirus GII type RNA-dependent RNA polymerase and capsid protein gene in excrement, vomit and other specimens of suspected Norovirus infected patients with diarrhea and vomiting as the main symptoms by disease prevention and control institutions at all levels, diarrhea virus detection network laboratories, sentinel hospitals and the like. Specifically, an HNB color-based reverse transcription loop-mediated isothermal amplification technology (RT-LAMP) is employed to analyze conserved sequences of Norovirus GII type RNA-dependent RNA polymerase and capsid protein gene by bioinformatical professional software, six primers are designed to identify a target sequence, and the six primers are completely matched with eight binding regions in the target sequence, so that an individual secondary amplification step of reverse transcription and nested PCR (polymerase chain reaction) can be omitted. Before reaction, an HNB (hydroxy naphthol blue) dye is added, and only one step is needed to finish 1h amplification in an isothermal environment of 65DEG C. Finally, visual inspection of color change and the white precipitate at a tube bottom is carried out so as to determine the detection result. Compared with traditional RT-PCR methods, the method involved in the invention has the advantages of better specificity, sensitivity and convenience.

Detection of GII type Norovirus by color-based reverse transcription loop-mediated isothermal amplification technology

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Color based loop-mediated isothermal amplification technology for detecting HPV6, HPV11, HPV16, HPV18, HPV52 and HPV58

Method for detecting CA16 virus gene by color-based reverse transcriptase loop-mediated isothermal amplification

Detection of GII type Norovirus by color-based reverse transcription loop-mediated isothermal amplification technology