CN102251061A - Nucleic acid dual fluorescence PCR (Polymerase Chain Reaction) detection kit for influenza A/B virus - Google Patents
Nucleic acid dual fluorescence PCR (Polymerase Chain Reaction) detection kit for influenza A/B virus Download PDFInfo
- Publication number
- CN102251061A CN102251061A CN2011102239136A CN201110223913A CN102251061A CN 102251061 A CN102251061 A CN 102251061A CN 2011102239136 A CN2011102239136 A CN 2011102239136A CN 201110223913 A CN201110223913 A CN 201110223913A CN 102251061 A CN102251061 A CN 102251061A
- Authority
- CN
- China
- Prior art keywords
- influenza
- virus
- seq
- probe
- enzyme
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Abstract
The invention provides a nucleic acid dual fluorescence PCR (Polymerase Chain Reaction) detection kit for an influenza A/B virus. The detection kit comprises an RNA (Ribonucleic Acid) enzyme inhibitor, an RT-PCR (Reverse Transcription-Polymerase Chain Reaction) liquid, an enzyme mixed liquor, an influenza A/B virus dual reaction liquid, a positive control and a negative control, wherein the influenza A/B virus dual reaction liquid comprises a component (1) which consists of a pair of primers for detecting the influenza A virus and a probe for detecting the influenza A virus and a component (2) which consists of a pair of primers for detecting the influenza B virus and a probe for detecting the influenza B virus; and the enzyme mixed liquor comprises a Taq enzyme and a reverse transcription enzyme. By adopting the detection kit, the influenza A and B viruses can be detected at the same time, and the problem that only one influenza virus can be detected in one reaction in the conventional product is solved. The detection kit has the advantages of easiness and convenience for operating, high repeatability, quick and objective detection result and the like, and has a great application prospect in the field of in-vitro diagnosis of influenza viruses.
Description
Technical field
The present invention relates to a kind of first type/Influenza B virus nucleic acid double fluorescent PCR detection kit that detects simultaneously, belong to biological technical field.
Background technology
Influenza is a kind of seasonal disease, and in the area, temperate zone, influenza can be popular in whole winter, and in the torrid areas, influenza virus all exists throughout the year, sees rainy season is popular more.Common influenza A is commonly called as cold, is to be the acute respiratory transmissible disease that influenza A virus causes by pathogenic agent, and main circulation way is the air droplet transmission.Influenza morbidity severity is relevant with the individual immunity situation.In general, only about 50% infected patient can develop into typical influenza clinical symptom.The influenza classical symptom is light with unexpected heating, dizziness headache, myalgia, constitutional symptom, simultaneously can be with having a sore throat and symptoms such as cough, nasal obstruction, runny nose, pectoralgia, ophthalmodynia, photophobia.Heating body temperature can reach 39～40 ℃, generally continues gradually to move back after 2～3 days.The clinical symptom of first type and second type influenza virus is similar, but the admission rate that influenza A virus causes is four times in Influenza B virus.Second type influenza virus has myositis and gastrointestinal symptom usually.
Influenza virus is that a kind of mankind of causing and animal suffer from grippal RNA viruses, belongs to Orthomyxoviridae family, and its viral genome constitutes (as Fig. 1) by 8 strand RNA sections.Different according to nucleoprotein (NP) with stromatin (M), be divided into first type, B-mode and third type.Influenza A virus is different according to surperficial hemagglutinin (HA) and neuraminidase (NA), is divided into 16 HA hypotypes and 9 NA hypotypes; B-mode and influenza virus C is regardless of hypotype.Influenza A virus is the easiest to morph, and flu outbreak is exactly that influenza A virus new subtype or old hypotype occur and reappears and cause; B-mode and influenza C is only relayed between the people, to break out and distribute the Flow Behavior feature in the part.
What cause flu outbreak mainly is influenza A virus.The detection method of influenza virus main virulent separation and Culture, immunology diagnosis and diagnosis of molecular biology.Many special methods have fast appearred in the ordinary method of the laboratory diagnosis that the separation and Culture of virus and immunology diagnosis are influenza in the diagnosis of molecular biology, three kinds of methods have following characteristics:
1) viral separation and Culture is higher to technical requirements, and expense is expensive, and length consuming time, and each laboratory separates positive rate and have nothing in common with each other can't satisfy the needs of handling great amount of samples during the viral prevalence simultaneously, only is used for experimental study at present.
Whether 2) serological method can not high-throughput be handled sample, can not accurately reflect currently to infect or carry virus.Also there is hysteresis quality in serological diagnosis, and time-consuming, the effort of this method can't satisfy requirement quick, early diagnosis.
3) viral nucleic acid detects: because above two kinds of methods are time-consuming, effort, in order to improve detection speed, can directly detect the nucleic acid of virus from clinical samples (throat swab, nose swab, nasopharynx or tracheae extract thing, phlegm).This method has advantages such as rapidity, accuracy and high sensitivity, can make early diagnosis to influenza infection, strong technical support is provided for the real-time analysis of epidemic situation and control.Real-time quantitative RT-PCR detection platform has wherein become the simplest and the most direct and reliable influenza virus detection method.
The product major part is taked the detection method at single virus at present, for the requirement of the multiple virus of clinical detection, must carry out the multitube detection to a sample and just can reach, and has increased detection cost and complex operation degree.
As seen from top, isolation of virus commonly used is more time-consuming, and there is hysteresis quality in serological diagnostic method, and the gene sequencing method is also comparatively complicated and loaded down with trivial details.
Summary of the invention
The objective of the invention is to overcome the deficiencies in the prior art part, a kind of first type/Influenza B virus nucleic acid double fluorescent PCR detection kit is provided.
First type of the present invention/Influenza B virus nucleic acid double fluorescent PCR detection kit comprises RNA enzyme inhibitors, RT-PCR reaction solution, enzyme mixed solution, first type/Influenza B virus double reaction liquid, positive control and negative control,
Wherein,
Said RNA enzyme inhibitors is diethylpyrocarbonate (DEPC) water;
Said RT-PCR reaction solution comprises 10 * damping fluid, MgCl2 and dNTPs;
Said first type/Influenza B virus double reaction liquid comprises following component:
Component (1): primer and a probe that detects influenza A virus by a pair of detection influenza A virus are formed; Wherein, the base sequence of two primers is respectively shown in SEQ ID No.1 and the SEQ ID No.2; The base sequence of probe is shown in the SEQ ID No.3, and 5 of this probe ' end is marked with the fluorescence report group, and 3 ' end is marked with the fluorescent quenching group;
Component (2): primer and a probe that detects Influenza B virus by a pair of detection Influenza B virus are formed; Wherein, the base sequence of two primers is respectively shown in SEQ ID No.4 and the SEQ ID No.5; The base sequence of probe is shown in the SEQ ID No.6, and 5 of this probe ' end is marked with the fluorescence report group, and 3 ' end is marked with the fluorescent quenching group;
Said enzyme mixed solution: comprise Taq enzyme and reversed transcriptive enzyme.
Table 1 is the sequence of above-mentioned first type/Influenza B virus primer and probe.
Preferably,
The primer of said detection influenza A virus and the proportioning of probe are respectively: SEQ ID No.1:SEQ ID No.2:SEQ ID No.3 is 600nM:600nM:200nM; The primer of said detection Influenza B virus and the proportioning of probe are respectively: SEQ ID No.4:SEQ ID No.5:SEQ ID No.6 is 400nM:400nM:400nM.
Said fluorescence report group is selected from FAM, HEX, ROX, CY3 or CY5 fluorescence report group; Described fluorescent quenching group is selected from BHQ1, BHQ2 or BHQ3 fluorescent quenching group.
Said reversed transcriptive enzyme is the M-MLV reversed transcriptive enzyme, and said Taq enzyme is a warm start Taq enzyme.
Said positive control is the virus-culturing fluid of deactivation; Said negative control is for removing RNA enzyme water.
Test kit of the present invention adopts the double fluorescent quantitative PCR technology, and design first type/Influenza B virus probe realizes detecting whether have first type and Influenza B virus simultaneously in same reaction system.The design of primer 5.0 special softwares is adopted in the design of primer and probe, and the primer and the probe of design are compared at the GeneBank of NCBI, detects the specificity of primer and probe.Primer and probe are synthetic in professional Synesis Company, adopt ultraviolet spectrophotometer to measure optical density value (A260nm/A280nm is qualified) between 1.8-2.0
What be most widely used in the at present domestic diagnostic nucleic acid is real-time fluorescence PCR technology based on fluorescence labeling probe.Detection probes is to comprise the go out oligonucleotide of group of 5 ' end reporter group and 3 ' end quenching.When probe is complete, because quenching group has greatly reduced the fluorescence that reporter group sends near reporter group.During primer extension, cut off by Taq enzyme (3 ' → 5 ' exonuclease activity) with template bonded probe, reporter group separates with quenching group, produces fluorescent signal.In each PCR circulation, all have new reporter group to be sheared, so the increase of fluorescence signal intensity is directly proportional with the quantity of amplified production.
The double fluorescent quantitative PCR technology then is to add two fluorescently-labeled probes of difference (at the probe mark FAM of target gene 1, at the probe mark HEX of target gene 2) in same reaction system.In the PCR reaction process, if sample to be checked comprises target gene 1, then the probe of flag F AM produces fluorescent signal; If sample to be checked comprises target gene 2, then the probe of mark HEX produces fluorescent signal.Like this, same tube reaction then can be determined two target genes (as shown in Figure 2).
First type of the present invention/Influenza B virus kit for detecting nucleic acid has following technique effect:
1) this test kit is easy and simple to handle and can effectively prevent to pollute, and the PCR fluoroscopic examination time (from sample disposal) only be 2-3 hour, and realizes in same reaction system whether detection exists first type or Influenza B virus simultaneously.The PCR fluoroscopic examination is a totally closed operation, adds the sample extract product and can no longer open the pipe lid afterwards, has reduced and has polluted the chance that produces.
2) can detect first type, Influenza B virus simultaneously, solve currently available products can only detect a kind of influenza virus in a pipe problem.The present invention also has advantages such as highly sensitive, that specificity good, repeatability is strong, detected result is quick and objective, has great application prospect in influenza virus in-vitro diagnosis field.
Description of drawings
Fig. 1 is the structure of influenza virus.
Fig. 2 is the double fluorescent quantitative PCR know-why.
Fig. 3 is the graphic representation that test kit of the present invention detects sample.
Fig. 4 is the sensitivity test result that test kit of the present invention detects influenza A virus.
Fig. 5 is the sensitivity test result that test kit of the present invention detects Influenza B virus.
Fig. 6 is the specificity test-results that test kit of the present invention detects influenza A virus.
Fig. 7 is the specificity test-results that test kit of the present invention detects Influenza B virus.
Embodiment
The first type of present embodiment/Influenza B virus nucleic acid double fluorescent quantitative PCR detection kit comprises RNA enzyme inhibitors, RT-PCR reaction solution, enzyme mixed solution, first type/Influenza B virus double reaction liquid, positive control and negative control,
Wherein, the RNA enzyme inhibitors is a DEPC water; The RT-PCR reaction solution comprises 10 * damping fluid, MgCl2 and dNTPs;
First type/Influenza B virus double reaction liquid comprises following component:
Component (1): primer and a probe that detects influenza A virus by a pair of detection influenza A virus are formed; Wherein, the base sequence of two primers is respectively shown in SEQ ID No.1 and the SEQ ID No.2; The base sequence of probe is shown in the SEQ ID No.3, and 5 of this probe ' end is marked with the fluorescence report group, and 3 ' end is marked with the fluorescent quenching group; Detecting the primer of influenza A virus and the proportioning of probe is respectively: SEQ ID No.1:SEQ ID No.2:SEQ ID No.3 is 600nM:600nM:200nM;
Component (2): primer and a probe that detects Influenza B virus by a pair of detection Influenza B virus are formed; Wherein, the base sequence of two primers is respectively shown in SEQ ID No.4 and the SEQ ID No.5; The base sequence of probe is shown in the SEQ ID No.6, and 5 of this probe ' end is marked with the fluorescence report group, and 3 ' end is marked with the fluorescent quenching group; Detecting the primer of Influenza B virus and the proportioning of probe is respectively: SEQ ID No.4:SEQ ID No.5:SEQ ID No.6 is 400nM:400nM:400nM.
The fluorescence report group is selected from FAM, HEX, ROX, CY3 or CY5 fluorescence report group; The fluorescent quenching group is selected from BHQ1, BHQ2 or BHQ3 fluorescent quenching group.
The enzyme mixed solution: comprise Taq enzyme and reversed transcriptive enzyme, reversed transcriptive enzyme is the M-MLV reversed transcriptive enzyme, and said Taq enzyme is a warm start Taq enzyme.
Positive control is the virus-culturing fluid of deactivation; Said negative control is for removing RNA enzyme water.
The using method of this test kit comprises the following steps:
1, extracts sample rna
1.1 get 200 ul influenza virus clinical samples, add 400 ul Binding Buffer supplemented with Poly (A), fully change high purifying strainer tube over to behind the mixing, centrifugal 15 s of 8000 rmp discard the waste liquid in the collection tube.
1.2 add 500ul Inhibitor Removal Buffer to strainer tube, centrifugal 1 min of 8000 rmp discards the waste liquid in the collection tube.
1.3 add 450ul Washing Buffer to strainer tube, centrifugal 1 min of 8000 rmp discards the waste liquid in the collection tube.
1.4 repeating step 1.3, high speed centrifugation 10 s then, this step must be removed waste liquid clean.
1.5 add 50ul Elution Buffer to strainer tube, room temperature leaves standstill 2 min, centrifugal 1 min of 8000 rmp, and centrifugal gained solution is purified RNA.
2 put into component first type/Influenza B virus detection reagent and RNA sample as shown in table 2 at each PCR reaction tubes, and wherein first type/Influenza B virus double reaction liquid is prepared according to table 3:
3, in the fluorescent quantitative PCR instrument, increase, carry out pcr amplification according to follow procedure:
4, after amplification finishes, judge whether to infect first type/Influenza B virus according to fluorescence curve, amplification curve as shown in Figure 3.
The result judges: fluorescence curve is " S " type curve and CT≤35.0 in the FAM passage, is judged as the influenza A virus positive; Do not have amplification of typical case's " S " type or CT〉35.0, be judged as the influenza A virus feminine gender.Fluorescence curve is " S " type curve and CT≤35.0 in the HEX passage, is judged as the Influenza B virus positive; Do not have amplification of typical case's " S " type or CT〉35.0, be judged as the Influenza B virus feminine gender.5 clinical sample concrete outcomes see Table 4.
The test of embodiment 2 susceptibilitys
Positive reference material is the virus-culturing fluid of deactivation, derives from Jiangsu Prov. Disease Preventing and Controlling Center.
Negative reference material is for removing RNA enzyme water, takes by weighing the DEPC of 1g with electronic balance, and the benefit purified water is to the 1000ml mixing, and 121 ℃/20 minutes high-temperature sterilizations in Autoclave are carried out mark, room temperature preservation then.
Adopt test kit of the present invention to detect.
Detected result shows that test kit of the present invention has good susceptibility, and the CT value reduces change (Fig. 4, Fig. 5) in gradient with concentration.Test-results shows that test kit of the present invention has the susceptibility of height for the diagnosis of first type/Influenza B virus.
The test of embodiment 3 specificitys
In order to detect the specificity of first type of the present invention/second influenza virus detection kit, detect respiratory syncytial virus, adenovirus hominis, human parainfluenza virus with first type of the present invention/Influenza B virus detection kit.
Detected result shows: the FAM passage only increases (Fig. 6) to influenza A virus, and the HEX passage is only to Influenza B virus increase (Fig. 7).Show that detection kit of the present invention can the specific amplification influenza virus, and not with other viral nucleic acid generation cross reaction.
Claims (5)
1. first type/Influenza B virus nucleic acid double fluorescent PCR detection kit is characterized in that, comprises RNA enzyme inhibitors, RT-PCR reaction solution, enzyme mixed solution, first type/Influenza B virus double reaction liquid, positive control and negative control,
Wherein,
Said RT-PCR reaction solution comprises 10 * damping fluid, MgCl2 and dNTPs;
Said first type/Influenza B virus double reaction liquid comprises following component:
Component (1): primer and a probe that detects influenza A virus by a pair of detection influenza A virus are formed; Wherein, the base sequence of two primers is respectively shown in SEQ ID No.1 and the SEQ ID No.2; The base sequence of probe is shown in the SEQ ID No.3, and 5 of this probe ' end is marked with the fluorescence report group, and 3 ' end is marked with the fluorescent quenching group;
Component (2): primer and a probe that detects Influenza B virus by a pair of detection Influenza B virus are formed; Wherein, the base sequence of two primers is respectively shown in SEQ ID No.4 and the SEQ ID No.5; The base sequence of probe is shown in the SEQ ID No.6, and 5 of this probe ' end is marked with the fluorescence report group, and 3 ' end is marked with the fluorescent quenching group;
Said enzyme mixed solution comprises Taq enzyme and reversed transcriptive enzyme.
2. first type according to claim 1/Influenza B virus nucleic acid double fluorescent PCR detection kit, it is characterized in that the primer of said detection influenza A virus and the proportioning of probe are respectively: SEQ ID No.1:SEQ ID No.2:SEQ ID No.3 is 600nM:600nM:200nM; The primer of said detection Influenza B virus and the proportioning of probe are respectively: SEQ ID No.4:SEQ ID No.5:SEQ ID No.6 is 400nM:400nM:400nM.
3. first type according to claim 1/Influenza B virus nucleic acid double fluorescent PCR detection kit is characterized in that said fluorescence report group is selected from FAM, HEX, ROX, CY3 or CY5 fluorescence report group; Described fluorescent quenching group is selected from BHQ1, BHQ2 or BHQ3 fluorescent quenching group.
4. first type according to claim 1/Influenza B virus nucleic acid double fluorescent PCR detection kit is characterized in that said reversed transcriptive enzyme is the M-MLV reversed transcriptive enzyme, and said Taq enzyme is a warm start Taq enzyme.
5. first type according to claim 1/Influenza B virus nucleic acid double fluorescent PCR detection kit is characterized in that said positive control is the virus-culturing fluid of deactivation; Said negative control is for removing RNA enzyme water.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011102239136A CN102251061A (en) | 2011-08-05 | 2011-08-05 | Nucleic acid dual fluorescence PCR (Polymerase Chain Reaction) detection kit for influenza A/B virus |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011102239136A CN102251061A (en) | 2011-08-05 | 2011-08-05 | Nucleic acid dual fluorescence PCR (Polymerase Chain Reaction) detection kit for influenza A/B virus |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN102251061A true CN102251061A (en) | 2011-11-23 |
Family
ID=44978629
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2011102239136A Pending CN102251061A (en) | 2011-08-05 | 2011-08-05 | Nucleic acid dual fluorescence PCR (Polymerase Chain Reaction) detection kit for influenza A/B virus |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102251061A (en) |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103614495A (en) * | 2013-12-02 | 2014-03-05 | 上海之江生物科技股份有限公司 | Joint detection kit for influenza viruses A and B and application thereof |
| CN108486282A (en) * | 2018-03-14 | 2018-09-04 | 南京岚煜生物科技有限公司 | It is a kind of to be used to detect A type, the kit of influenza B virus and its application method |
| CN108504775A (en) * | 2018-03-14 | 2018-09-07 | 南京岚煜生物科技有限公司 | A kind of detection A type, the kit of influenza B virus and its application method based on micro-fluidic chip |
| CN111254228A (en) * | 2020-05-06 | 2020-06-09 | 上海思路迪医学检验所有限公司 | Kit for detecting novel coronavirus and influenza virus |
| CN111411172A (en) * | 2020-02-28 | 2020-07-14 | 江苏硕世生物科技股份有限公司 | Probe and primer composition for simultaneously detecting novel human coronavirus, influenza A virus and influenza B virus |
| CN113637795A (en) * | 2020-04-27 | 2021-11-12 | 上海星耀医学科技发展有限公司 | Detection method and kit for influenza A/B virus and novel coronavirus |
| WO2023077490A1 (en) * | 2021-11-06 | 2023-05-11 | 江汉大学 | Combination of mnp markers of influenza a, b and c viruses, primer pair combination, kit, and uses of combination, primer pair combination and kit |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101921870A (en) * | 2009-06-17 | 2010-12-22 | 上海人类基因组研究中心 | Nucleotide sequence, method and kit for detecting human influenza A virus |
| CN101942525A (en) * | 2010-05-06 | 2011-01-12 | 广州市华南医学病毒学研究所 | One-tube method with multiplex detection for human Influenza A and B and new Influenza A H1N1 virus and kit |
-
2011
- 2011-08-05 CN CN2011102239136A patent/CN102251061A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101921870A (en) * | 2009-06-17 | 2010-12-22 | 上海人类基因组研究中心 | Nucleotide sequence, method and kit for detecting human influenza A virus |
| CN101942525A (en) * | 2010-05-06 | 2011-01-12 | 广州市华南医学病毒学研究所 | One-tube method with multiplex detection for human Influenza A and B and new Influenza A H1N1 virus and kit |
Non-Patent Citations (1)
| Title |
|---|
| 徐昌平等: "双重荧光定量RT-PCR快速检测A、B型流感病毒的研究", 《中国卫生检验杂志》 * |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103614495A (en) * | 2013-12-02 | 2014-03-05 | 上海之江生物科技股份有限公司 | Joint detection kit for influenza viruses A and B and application thereof |
| CN103614495B (en) * | 2013-12-02 | 2014-12-10 | 上海之江生物科技股份有限公司 | Joint detection kit for influenza viruses A and B and application thereof |
| CN108486282A (en) * | 2018-03-14 | 2018-09-04 | 南京岚煜生物科技有限公司 | It is a kind of to be used to detect A type, the kit of influenza B virus and its application method |
| CN108504775A (en) * | 2018-03-14 | 2018-09-07 | 南京岚煜生物科技有限公司 | A kind of detection A type, the kit of influenza B virus and its application method based on micro-fluidic chip |
| CN111411172A (en) * | 2020-02-28 | 2020-07-14 | 江苏硕世生物科技股份有限公司 | Probe and primer composition for simultaneously detecting novel human coronavirus, influenza A virus and influenza B virus |
| CN113637795A (en) * | 2020-04-27 | 2021-11-12 | 上海星耀医学科技发展有限公司 | Detection method and kit for influenza A/B virus and novel coronavirus |
| CN111254228A (en) * | 2020-05-06 | 2020-06-09 | 上海思路迪医学检验所有限公司 | Kit for detecting novel coronavirus and influenza virus |
| CN111254228B (en) * | 2020-05-06 | 2020-09-11 | 上海思路迪医学检验所有限公司 | Kit for detecting novel coronavirus and influenza virus |
| WO2023077490A1 (en) * | 2021-11-06 | 2023-05-11 | 江汉大学 | Combination of mnp markers of influenza a, b and c viruses, primer pair combination, kit, and uses of combination, primer pair combination and kit |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Mahony et al. | Multiplex loop-mediated isothermal amplification (M-LAMP) assay for the detection of influenza A/H1, A/H3 and influenza B can provide a specimen-to-result diagnosis in 40 min with single genome copy sensitivity | |
| Beck et al. | Molecular diagnosis of respiratory viruses | |
| JP6329370B2 (en) | Simultaneous diagnosis kit for diseases caused by respiratory viruses | |
| CN102251061A (en) | Nucleic acid dual fluorescence PCR (Polymerase Chain Reaction) detection kit for influenza A/B virus | |
| Ieven | Currently used nucleic acid amplification tests for the detection of viruses and atypicals in acute respiratory infections | |
| CN102286639A (en) | Type A H1N1/influenza A virus nucleic acid dual fluorescent polymerase chain reaction (PCR) detection kit | |
| CN102337351B (en) | Typing detection kit for influenza virus | |
| CN105296673A (en) | Influenza A virus molecular detection kit and preparation method thereof | |
| CN111206121A (en) | Kit for detecting novel coronavirus orflab and S genes | |
| Li et al. | A two-tube multiplex reverse transcription PCR assay for simultaneous detection of sixteen human respiratory virus types/subtypes | |
| CN103233082B (en) | Kit for testing H7N9 bird flu virus using real time fluorescence quantitative PCR | |
| TW202018093A (en) | Method for influenza a virus and influenza b virus detection | |
| CN103789455B (en) | Group of real-time fluorescent QRT-PCR primers capable of detecting cattle, sheep akabane virus by one-step method, probe and usage method thereof | |
| CN104846121A (en) | Virus triple fluorescence quantitative RT-PCR detection method and virus triple fluorescence quantitative RT-PCR detection kit | |
| US10329630B2 (en) | Compositions and methods for detection and discrimination of emerging influenza virus subtypes | |
| CN110343784A (en) | The composition and kit of quadruple influenza nucleic acids detection based on melting curve | |
| EP3060686B1 (en) | Compositions and methods for detection of influenza viruses | |
| Matsuda et al. | A novel high-speed droplet-polymerase chain reaction can detect human influenza virus in less than 30 min | |
| GB2401175A (en) | Detection of SARS virus by PCR | |
| CN103409554B (en) | Nucleic acid detection kit for rapidly detecting measles virus/rubella virus | |
| CN102146485B (en) | One-step RT-PCR (Reverse Transcription-Polymerase Chain Reaction) detection kit for influenza virus | |
| CN104593524A (en) | Nucleic acid detection kit for rapidly detecting respiratory syncytial virus A and B and application thereof | |
| CN112410465A (en) | Novel coronavirus SARS-CoV-2ORF1ab and N gene constant temperature amplification primer group and kit | |
| CN105296675A (en) | Nucleic acid detecting kit for influenza B viruses and preparation method of nucleic acid detecting kit | |
| WO2023109031A1 (en) | Respiratory pathogen detection kit, and preparation method therefor and use thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C12 | Rejection of a patent application after its publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20111123 |