CN102337351A - Typing detection kit for influenza virus - Google Patents
Typing detection kit for influenza virus Download PDFInfo
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- CN102337351A CN102337351A CN2010102294961A CN201010229496A CN102337351A CN 102337351 A CN102337351 A CN 102337351A CN 2010102294961 A CN2010102294961 A CN 2010102294961A CN 201010229496 A CN201010229496 A CN 201010229496A CN 102337351 A CN102337351 A CN 102337351A
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Abstract
The invention relates to a typing detection kit for an influenza virus, especially to a kit for detecting influenza virus A, influenza virus B and influenza virus A (H1N1) with a multiple real-time fluorescent polymerase chain reaction technology. The kit mainly comprises an RT-PCR (real-time polymerase chain reaction) reaction solution, a mixed solution of primer and probe, RT-PCR reaction enzymes, DEPC H2O, and packing boxes for separation and centralized packaging of the reagent bottles or tubes. Able to distinguish influenza virus A, influenza virus B and influenza virus A (H1N1) accurately, the kit of the invention can be widely applied in multiple fields like differential diagnosis and epidemic prevention and control of influenza caused by the above influenza viruses.
Description
Technical field
The present invention relates to the test kit that a kind of influenza virus somatotype detects, particularly relate to a kind of test kit that utilizes the multiple real time fluorescence polymerase chain reaction technology to detect influenza A virus, Influenza B virus and H1N1virus.This test kit can accurately be distinguished first type, Influenza B virus and H1N1virus, can be widely used in by a plurality of fields such as the differential diagnosis of the caused influenza of above-mentioned influenza virus, epidemic situation prevention and control.
Background technology
Seasonal influenza is a kind of common acute respiratory transmissible disease that is caused by influenza virus.North China, how occurred frequently in the winter time seasonal influenza is; And at southern area, the four seasons all have case to take place, and epidemic peak can appear in summer and winter.Human to the general susceptible of influenza virus.Influenza virus also has the people because touch the article (like tableware, teacup or toy etc.) that polluted influenza virus mainly through the infected cough or propagations of sneezing, and contacts mouth or nose and the infective virus of oneself then again.Be about 1～3 day the latent period of seasonal influenza, and Acute onset shows as high heat, headache, myalgia, toxicity symptom such as weak usually, can be with symptoms such as pharyngalgia, runny nose, dry cough, gastrointestinal upsets.Main complication is bacterial pneumonia, ear infection, sinus infection, dehydration, old man, children, with severe complications takes place behind some basic disease or the person's of the having a delicate constitution influenza virus infection easily, in addition dead.Though can obtain certain immunizing power after being ill, because variation by a small margin takes place through regular meeting in influenza virus sub-strain inside, i.e. " antigenic drift ", the people is to the virus after making a variation still susceptible.Flu outbreak refers to the influenza of global range and breaks out.It can be brand-new causing pandemic influenza virus, also can be that old influenza virus reappears.Almost do not resist the immunizing power of this virus in the human body, still do not have effective vaccine again.Influenza virus can be propagated in the crowd fast, and serious disease can take place the infected, even dead.Disease can involve the whole country and even the whole world at short notice.
Influenza virus belongs to orthomyxoviridae family (Orthomyxoviridae) on viral taxonomy.Its genome is segmented (first, B-mode strain contain 8 sections, and third type only contains 7 sections, a few proteic sections of coding neuraminidase), the RNA of sub-thread minus strand.First, B-mode strain genome encode 10 and 11 kind of albumen respectively.Because genome is segmented, so be prone to produce not gene resortment between homophyletic of homotype.Influenza virus; Especially the constant origination point sudden change of influenza virus A hominis HA gene; Cause the HA protein molecular upper amino acid sequence of its coding to be replaced, cause constant the drifting about of its antigenicity, each antigenic drift often brings influenza pandemic in various degree.Influenza A virus can be divided into many hypotypes again according to its surperficial hemagglutinin (H) and neuraminidase (N) protein structure and genetic characteristics thereof; So far the blood clotting found of influenza A virus have 15 hypotypes (H1-H15), and neuraminidase has 9 hypotypes (N1-N9).H1N1virus is that influenza A virus belongs to (Influenza virus A), and typical virion is spherical, and diameter is 80nm-120nm, and cyst membrane is arranged.The projection gp that many radial arrangement are arranged on the cyst membrane is respectively red corpuscle hemagglutinin (HA), neuraminidase (NA) and stromatin M2.In the virion is nucleocapsid, and shape is symmetrical in the shape of a spiral, and diameter is 10nm, is the sub-thread minus-stranded rna virus, and genome is about 13.6kb, is made up of 8 independent segments that differ in size.Virus is responsive to common disinfectantses such as ethanol, Iodophor, the tincture of iodine; To thermo-responsive, but the deactivation in following 30 minutes of 56 ℃ of conditions.On March 18th, 2009; Mexico finds the H 1 N 1 influenza A virus infection case successively; And several deaths appear, and the Influenza A H1N1 epidemic situation occurs in a big way in the whole world subsequently, and the World Health Organization was also once being carried the flu outbreak warning level to 6 grades.China has included Influenza A H1N1 in the Category B notifiable disease of " the People's Republic of China's law on the prevention and control of infectious diseases " regulation, and takes the preventive and control measure of category A infectious disease.After 2010, the influenza epidemic situation is stepped into the mitigation stage.
The general method of influenza virus of identifying mainly contains following several method: (1) viral isolation identification.Adopting chicken embryo and/or mdck cell to carry out influenza virus separates.Sample is processed suspension inoculation in 10～11 age in days chick embryo allantoic cavities, collects allantoic fluid behind the 48h, can cultivate to gather in the crops virus-culturing fluid or to carry out titre and identify with mdck cell again.This method detection time is long and the operator required than higher; (2) serological method also is the conventional means that virus is identified, judges the kind and the hypotype of virus through serological identification.Disturb but often receive aspects such as nonspecific agglutination factor, antibodies specific in the experimentation, aspects such as antigenic stdn, Monoclonal Antibody technology are proposed stricter requirement.(3) molecular Biological Detection method.Protocols in Molecular Biology has been widely used in the early stage quick diagnosis of influenza.Rt-polymerase chain reaction is that RT-PCR has high degree of specificity and susceptibility; Be to develop the most sophisticated molecular diagnostic techniques at present; Can detect Influenza Virus RNA from gene level; Shortened the detection time of cause of disease greatly, for the diagnosis of influenza virus provides more responsive, method faster.Present most of laboratory all adopts the polymerase chain reaction to carry out the evaluation of virus.Influenza virus is single stranded RNA, and therefore suitable employing reverse transcriptase polymerase chain reaction carries out the amplification of viral nucleic acid.
The real-time fluorescence PCR technology is a kind of direct detection nucleic acid and the Protocols in Molecular Biology that carries out nucleic acid quantification, has highly sensitively, and specificity is good, and is simple, advantages such as cheapness.It adds fluorophor a kind of in the PCR reaction system, utilize fluorescent signal accumulation monitoring PCR reaction process in real time, through the result data analysis starting template is carried out quantitatively.The present invention adopts the multiple real time fluorescence PCR method in the real-time fluorescence PCR technology; Three kinds of influenza viruses (influenza A virus, Influenza B virus, H1N1virus) nucleic acid is increased; According to the amplification situation of the three fluorescence of mark, differentiate three kinds of influenza viruses.The influenza that test kit of the present invention can apply to caused by influenza A virus, Influenza B virus and H1N1virus is widely monitored.
Summary of the invention
The object of the present invention is to provide a kind of multiple real time fluorescence polymerase chain reaction technology that utilizes to carry out the test kit that the influenza virus somatotype detects, utilize this test kit can accurately distinguish influenza A virus, Influenza B virus and H1N1virus.
On the basis that the nucleotide sequence of all known influenza A viruss, Influenza B virus and H1N1virus strain is compared on to GENBANK; Seek the specificity conserved regions of three kinds of nucleic acid sequences respectively, and to conserved regions design target nucleotide primer and probe.These primer probes are containing hot resistant DNA polymerase, the RT-PCR reaction enzymes of reversed transcriptive enzyme, high quality deoxyribonucleoside triphosphate (dNTPs) system and contain Mg
2+In the RT-PCR reaction solution Deng composition, realize the cyclic amplification of external nucleic acid through the fluorescent PCR appearance.
Test kit involved in the present invention mainly comprises: 1) RT-PCR reaction solution, primer probe mixed solution, RT-PCR reaction enzymes system, DEPC H
2O and 2) packing box of separation and concentrated these reagent bottles of packing or pipe.
A preferred embodiment of the present invention is that primer probe mixed solution is by the specific forward and reverse primer of a pair of influenza A virus;, the specific probe of influenza A virus; The specific forward and reverse primer of a pair of Influenza B virus;, the specific probe of Influenza B virus; The forward and reverse primer of a pair of influenza A H 1 N 1 virus specific; Article one, the probe of influenza A H 1 N 1 virus specific is formed, and the sequence that it is characterized in that specific forward of influenza A virus and reverse primer is respectively 5 '-TAAAGATGAGTCTTCTAACCGAG-3 ' (SEQ ID NO:1) and 5 '-CAAGATCTGTGTTCTTTCCTGC-3 ' (SEQ ID NO:2); The sequence of the specific probe of influenza A virus is 5 '-CGAAACGTACGTTCTTTCTATCATCCC-3 ' (SEQ ID NO:3), and the two ends of probe are combined with fluorescence generation group FAM and fluorescent quenching group B HQ1 respectively; The sequence of specific forward of Influenza B virus and reverse primer is respectively 5 '-GTTGGTAAACGGAACATTCCTCAAAC-3 ' (SEQ ID NO:4) and 5 '-GCAACAAGCCTTCCACTCTGGTC-3 ' (SEQ ID NO:5); The sequence of the specific probe of Influenza B virus is 5 '-CCCAATGGATACAAGTCCTTATCAACTC-3 ' (SEQ ID NO:6), and the two ends of probe are combined with fluorescence generation group HEX and fluorescent quenching group Eclipse respectively; The forward of influenza A H 1 N 1 virus specific and the sequence of reverse primer are respectively 5 '-GGCCATTGCCGGTTTCA-3 ' (SEQ ID NO:7) and 5 '-TTGTTAGTAATCTCGTCAATGGCATT-3 ' (SEQ ID NO:8); The sequence of the probe of influenza A H 1 N 1 virus specific is 5 '-ATATGCAGCCGACCTGAAGAGCACACA-3 ' (SEQ ID NO:9), and the two ends of probe are combined with fluorescence generation group Cy5 and fluorescent quenching group Eclipse respectively.
Another preferred embodiment of the present invention is that primer concentration is 0.2mmol/L in the primer probe mixed solution, and concentration and probe concentration is 0.1mmol/L.
Another preferred embodiment of the present invention be the RT-PCR reaction solution by Tris-HCl (50mmol/L, pH8.0), MgCl
2(8mmol/L), KCl (250mmol/L) forms.
Another preferred embodiment of the present invention is that RT-PCR reaction enzymes system is made up of warm start Taq enzyme, reversed transcriptive enzyme, dNTPs.Reversed transcriptive enzyme can be selected reversed transcriptive enzymes commonly used such as mMLV.Warm start Taq enzyme, reversed transcriptive enzyme, dNTPs all can adopt the commercially available prod, and like the product of Qiagen company, wherein the consumption of warm start Taq enzyme is 5U in everyone part RT-PCR reaction enzymes system, and the reversed transcriptive enzyme consumption is 10U, and the dNTPs consumption is 10mmol.
The condition that another preferred embodiment of the present invention is a pcr amplification is: 50 ℃ 15 minutes, 95 ℃ 15 minutes; 94 ℃ 15 seconds, 58 ℃ 45 seconds, 40 circulations (58 ℃ time collect fluorescent signal).
A preferred embodiment of the present invention is that test kit provides extra quality control product, is respectively negative quality control product and positive quality control product.Negative quality control product is a saline water; Positive quality control product is the virus-culturing fluid of deactivation; Form by influenza A virus and two kinds of viruses of Influenza B virus; Virus strain is available from ATCC, and wherein influenza A virus can be selected one of following numbering: VR-1284, VR-1285, VR-1287, VR-1289, VR-1469, VR-1520, VR-1608, VR-1609, VR-1641, VR-1642, VR-1679, VR-1680, VR-1682, VR-1683, VR-1737, VR-219, VR-333, VR-544, VR-546, VR-547, VR-776, VR-777, VR-810, VR-822, VR-825, VR-897, VR-95, VR-96, VR-97, VR-98, VR-99.Influenza B virus can be selected one of following numbering: VR-101, VR-102, VR-103, VR-1535, VR-1735, VR-295, VR-296, VR-523, VR-786, VR-787, VR-788, VR-789, VR-790.Two kinds of virus concentrations are 5.0 * 10
3～5.0 * 10
6Between the PFU/mL, preferred 5.0 * 10
3PFU/mL, and mix at 1: 1 with volume ratio and to be prepared into positive quality control product.Carry out sample detection to be checked in the test kit and can carry out the detection of two special quality control article simultaneously; Only FAM, HEX and CY5 passage fluorescent signal all are positive when positive quality control product detects; When three passage fluorescent signals of negative quality control product detection all were negative, the detected result of sample to be checked was just effective.
The test kit of the present invention sample to be checked that increases is accomplished by commercially available quantitative real time PCR Instrument automatically, and is simple to operate, consuming time few, and reduced the generation of polluting to greatest extent.Detected result can be used for a plurality of area researches such as influenza virus somatotype, auxiliary clinical diagnosis and routine monitoring.
The present invention compared with prior art; Advantage is: 1. influenza virus nucleic acid amplification level is detected; Can reflect the state of first type in patient's body, B-mode and H 1 N 1 influenza A virus infection, can be used for the monitoring and the control of influenza, help the early diagnosis and therapy of disease; 2. the multiple real time fluorescence round pcr has higher flux to multiple viral nucleic acid specificity property sequences Design primer, probe, and easy and simple to handle, the advantage that reduces cost relatively is applicable to that more Most patients detects; Whether 3. a sample one-time detection can be distinguished is the type of influenza infection and infection, has improved detection efficiency greatly.
Description of drawings
Fig. 1 shows the reaction conditions of pcr amplification.
The amplification curve of the negative quality control product of Fig. 2 visualizingre agent box.Do not have S type amplification curve among the figure, explain that FAM, HEX and CY5 passage fluorescent signal all are negative.
The amplification curve of Fig. 3 visualizingre agent box positive quality control product.The amplification curve of FAM passage is the S type among the figure, explains that the fluorescent signal of this passage is positive.
The amplification curve of Fig. 4 visualizingre agent box positive quality control product.The amplification curve of HEX passage is the S type among the figure, explains that the fluorescent signal of this passage is positive.
The amplification curve of Fig. 5 visualizingre agent box positive quality control product.The amplification curve of CY5 passage is the S type among the figure, explains that the fluorescent signal of this passage is positive.
Fig. 6 shows the amplification curve of 3 routine negatives.Do not have S type amplification curve among the figure, explain to detect and do not have three kinds of influenza nucleic acids in the sample.
Fig. 7 shows the amplification curve of 1 routine positive throat swab sample.The amplification curve of FAM passage is the S type among the figure, explains that this sample has the amplification of influenza A virus nucleic acid, has influenza A virus in the expression sample.
Fig. 8 shows the amplification curve of 1 routine positive throat swab sample.The amplification curve of HEX passage is the S type among the figure, explains that this sample has the amplification of Influenza B virus nucleic acid, has Influenza B virus in the expression sample.
Fig. 9 shows the amplification curve of 1 routine positive throat swab sample.The amplification curve of FAM passage and Cy5 passage is the S type among the figure, explains that this sample has the amplification of H1N1virus nucleic acid, has H1N1virus in the expression sample.
Embodiment
Below in conjunction with specific embodiment, further set forth the present invention.Should be understood that these embodiment only to be used to the present invention is described and be not used in the restriction scope of the present invention.
1, preparation comprises the test kit of following moity: primer probe mixed solution (25 μ l/ pipe) 1 pipe; RT-PCR reaction solution (250 μ l/ pipe), RT-PCR reaction enzymes system (75 μ l/ pipe) 1 pipe, positive quality control product (200 μ l/ pipe) 1 pipe; Negative quality control product (200 μ l/ pipe) 1 pipe, DEPC H
2O (2000 μ l/ pipe) 1 pipe.
2, the collection of sample, storage and transport
2.1 sample type: nose swab, throat swab, nasopharynx are drawn respiratory tract samples such as thing.
2.2 collection of specimens, preserve and transport: get nasal cavity or throat secretory product with the disinfecting silk or cotton swab; And input is equipped with in the small test tube of sterile saline or Eagle liquid or 0.5% lactoalbumin hydrolysate Hanks liquid 2mL immediately; Post label, put take to the laboratory in the curling stone or low temperature (20 ℃～-70 ℃) frozen.The long-distance employing dry ice that transports of sample.
3, nucleic acid extraction:
Can adopt commercially available RNA to extract test kit, suggestion uses QIAamp Viral RNA Mini Kit to extract test kit, and operates according to the specification sheets of test kit, collects 50 μ l RNA solution at last, directly detects or be stored in-20 ℃.
4, real-time fluorescence quantitative PCR amplification and detection
4.1 reagent is prepared: primer probe mixed solution, RT-PCR reaction solution, the RT-PCR reaction enzymes of getting respective amount in proportion are that (primer probe mixed solution 1 μ l/ person-portion+RT-PCR reaction solution 10 μ l/ person-portion+RT-PCR enzymes are 3 μ l/ person-portion+DEPC H
2O 16 μ l/ person-portions), fully be distributed into the PCR reaction tubes by 30 μ l/ pipe behind the mixing, subsequent use.
4.2 application of sample: in the PCR reaction tubes, add positive and negative quality control product, sample rna solution 20 μ l after extracting respectively, the tight pipe lid of lid is put into the instrument sample cell.
4.3 editor: (ABI Prism 7500 quantitative real time PCR Instruments)
Open the Setup window, positive and negative quality control product and sample to be measured are set in proper order, and the sample title is set in the Name hurdle by correspondence.Choose all that sample well is set, double-click, select Add Detector, selecting Reporter is that FAM and Quencher are BHQ1, and selecting Reporter again is that HEX and Quencher are Eclipse; Selecting Reporter then is that Cy5 and Quencher close window behind the Eclipse.In Passive Reference, select (none).Open the instrument window cycling condition be set: 50 ℃ 15 minutes, 95 ℃ 15 minutes; 94 ℃ 15 seconds, 58 ℃ 45 seconds, 40 circulations (seeing accompanying drawing 1).All preserve file after being provided with and accomplishing, operation.
4.4 interpretation of result:
Reaction finishes the back and preserves the detection data file.Under Results, open Amp plot window.Select the purpose sample position of analysis.Change Baseline numerical value into start:3, stop:10, and open manual and set Threshold:1.5 ± 100000.Numerical value is opened Graph settings window on the double-click Rn coordinate, changes Log among the Post Run Settings into Linear, opens Analysis preferences window behind the OK, under the Analysis menu, selects the automatic analytical results of Analyze.
Choose 3 examples and be accredited as the influenza virus feminine gender through the virus culture method; 3 examples are accredited as influenza virus male sample through the virus culture method, the nucleic acid extraction of sample, and pcr amplification and interpretation of result step are carried out with reference to embodiment 1; Carry out the moon simultaneously, the detection of positive quality control product.
Detected result: the amplification curve of negative quality control product not S-type (seeing accompanying drawing 2); The amplification curve of positive quality control product is an obvious S type curve (seeing accompanying drawing 3,4,5); The positive and negative quality control product all meets the Quality Control requirement of test kit, and therefore the detected result of sample to be checked is effective.But all detect the amplification (seeing accompanying drawing 6,7,8,9) of corresponding first type, B-mode and H1N1virus nucleic acid by the detected result knowledge capital test kit of sample to be checked
Detected result and the virus culture qualification result of 6 routine samples fits like a glove in this test, explain that it is feasible utilizing the detection of this test kit and differentiation first type, B-mode and H1N1virus.This test kit is easy and simple to handle, and detection time is short, can realize high throughput testing, and it is cheap in addition, is expected to be applied to the monitoring of the clinical detection and the seasonal influenza of influenza.
Claims (6)
1. an influenza virus parting detecting reagent comprises: 1) RT-PCR reaction solution, primer probe mixed solution, RT-PCR reaction enzymes system, DEPC H
2O and 2) packing box of separation and concentrated these reagent bottles of packing or pipe, wherein primer probe mixed solution is by the specific forward and reverse primer of a pair of influenza A virus;, the specific probe of influenza A virus; The specific forward and reverse primer of a pair of Influenza B virus, the specific probe of Influenza B virus, the forward and reverse primer of a pair of influenza A H 1 N 1 virus specific; Article one, the probe of influenza A H 1 N 1 virus specific is formed, and it is characterized in that:
The sequence of specific forward of influenza A virus and reverse primer is respectively 5 '-TAAAGATGAGTCTTCTAACCGAG-3 ' and 5 '-CAAGATCTGTGTTCTTTCCTGC-3 '; The sequence of the specific probe of influenza A virus is 5 '-CGAAACGTACGTTCTTTCTATCATCCC-3 ', and the two ends of probe are combined with fluorescence generation group FAM and fluorescent quenching group B HQ1 respectively;
The sequence of specific forward of Influenza B virus and reverse primer is respectively 5 '-GTTGGTAAACGGAACATTCCTCAAAC-3 '; With 5 '-GCAACAAGCCTTCCACTCTGGTC-3 '; The sequence of the specific probe of Influenza B virus is 5 '-CCCAATGGATACAAGTCCTTATCAACTC-3 ', and the two ends of probe are combined with fluorescence generation group HEX and fluorescent quenching group Eclipse respectively;
The forward of influenza A H 1 N 1 virus specific and the sequence of reverse primer are respectively 5 '-GGCCATTGCCGGTTTCA-3 ' and 5 '-TTGTTAGTAATCTCGTCAATGGCATT-3 '; The sequence of the probe of influenza A H 1 N 1 virus specific is 5 '-ATATGCAGCCGACCTGAAGAGCACACA-3 ', and the two ends of probe are combined with fluorescence generation group Cy5 and fluorescent quenching group Eclipse respectively.
2. test kit according to claim 1, its characteristic are that also primer concentration is 0.2mmol/L in the primer probe mixed solution, and concentration and probe concentration is 0.1mmol/L.
3. test kit according to claim 1, its characteristic are that also the RT-PCR reaction solution is 8.0 50mmol/LTris-HCl, 8mmol/L MgCl by the pH value
2, 250mmol/L KCl forms.
4. test kit according to claim 1; Wherein RT-PCR reaction enzymes system is made up of warm start Taq enzyme, reversed transcriptive enzyme, dNTPs; The consumption that is characterised in that warm start Taq enzyme in everyone part RT-PCR reaction enzymes system is 5U, and the reversed transcriptive enzyme consumption is 10U, and dNTPs is 10mmol.
5. according to the test kit of claim 1, its characteristic is that also the condition of pcr amplification is: 50 ℃ 15 minutes, 95 ℃ 15 minutes; 94 ℃ 15 seconds, 58 ℃ 45 seconds, 45 circulations.
6. according to the test kit of claim 1, negative quality control product and positive quality control product are provided also, have it is characterized in that negative quality control product is a saline water, positive quality control product is the virus-culturing fluid of deactivation, is 5.0 * 10 by concentration
3PFU/mL influenza A virus and Influenza B virus mixed in 1: 1 by volume.
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| CN104032034A (en) * | 2014-03-07 | 2014-09-10 | 王全意 | Method for simultaneously detecting influenza A virus, influenza B virus and influenza C virus and kit |
| CN104388587A (en) * | 2014-11-03 | 2015-03-04 | 深圳市生科源技术有限公司 | One-step detection kit for influenza B virus and influenza B virus detection method |
| CN105950775A (en) * | 2016-07-08 | 2016-09-21 | 中山大学达安基因股份有限公司 | Kit for detecting NPM (Nucleophosmin)1 gene mutation types |
| CN106854682A (en) * | 2016-12-27 | 2017-06-16 | 上海派森诺生物科技股份有限公司 | A kind of method that parting is carried out using gene infected by influenza |
| CN107034310A (en) * | 2017-01-24 | 2017-08-11 | 南方医科大学 | It is a kind of while detecting the primer sets of a variety of influenza viruses, probe groups and its kit |
| CN107287346A (en) * | 2016-03-30 | 2017-10-24 | 广东省疾病预防控制中心 | Multiple fluorescence quantitative PCR detection kit and its application for avian influenza virus N subtype gene partings |
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| CN104032034A (en) * | 2014-03-07 | 2014-09-10 | 王全意 | Method for simultaneously detecting influenza A virus, influenza B virus and influenza C virus and kit |
| CN104388587A (en) * | 2014-11-03 | 2015-03-04 | 深圳市生科源技术有限公司 | One-step detection kit for influenza B virus and influenza B virus detection method |
| CN107287346A (en) * | 2016-03-30 | 2017-10-24 | 广东省疾病预防控制中心 | Multiple fluorescence quantitative PCR detection kit and its application for avian influenza virus N subtype gene partings |
| CN107287346B (en) * | 2016-03-30 | 2020-11-06 | 广东省疾病预防控制中心 | Multiplex fluorescence quantitative PCR detection kit for avian influenza virus N subtype genotyping and application thereof |
| CN105950775A (en) * | 2016-07-08 | 2016-09-21 | 中山大学达安基因股份有限公司 | Kit for detecting NPM (Nucleophosmin)1 gene mutation types |
| CN105950775B (en) * | 2016-07-08 | 2019-06-11 | 中山大学达安基因股份有限公司 | A kind of kit detecting NPM1 gene mutation typing |
| CN106854682A (en) * | 2016-12-27 | 2017-06-16 | 上海派森诺生物科技股份有限公司 | A kind of method that parting is carried out using gene infected by influenza |
| CN107034310A (en) * | 2017-01-24 | 2017-08-11 | 南方医科大学 | It is a kind of while detecting the primer sets of a variety of influenza viruses, probe groups and its kit |
| CN107893130A (en) * | 2017-12-26 | 2018-04-10 | 湖南圣湘生物科技有限公司 | The application method of the primer and probe of influenza virus parting detection, kit and kit |
| CN113249519A (en) * | 2021-04-21 | 2021-08-13 | 深圳市儿童医院 | Adenovirus detection typing kit and method for peripheral blood sample |
| CN113278735A (en) * | 2021-05-26 | 2021-08-20 | 首都医科大学附属北京朝阳医院 | Human respiratory virus nucleic acid multiplex detection primer, probe and kit |
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