CN103981282A - Method for collecting, preserving and detecting mosquito-borne viruses under normal temperature - Google Patents
Method for collecting, preserving and detecting mosquito-borne viruses under normal temperature Download PDFInfo
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Abstract
The invention relates to a method for collecting, preserving and detecting mosquito-borne viruses under normal temperature, which belongs to the collection, preservation and detection of virus. According to the invention, biological characteristics of virus discharged due to saliva spit during a process that mosquito sucks sugar solution are used, the captured mosquito is placed in a penicillin bottle, voile cages with quantity adapting to mosquito is designed, the sugar solution with certain concentration and RNA preservation liquid are added drop by drop on a piece of filter paper, filter paper is taken as a virus RNA carrier under normal temperature to collect virus, laboratory detection on the virus is carried out in 7 days of carrier preservation under normal temperature, analysis is accurate, the problems of field transport, carrying and preservation by a cold chain technology are prevented, virus collection and preservation efficiency can be increased, and the manpower and material resources can be saved.
Description
Technical field
The invention belongs to viral collection, preservation and detection, particularly carapuru virus is collected, is preserved and detects.
Background technology
In the pathogenic micro-organism that Mosquito Vectors carries, virus is that cause serious disease main causes a disease thereby extensively concerned, as encephalitis b virus, dengue fever virus etc.Mosquito monitoring is an aspect of the sick monitoring of mosquito matchmaker just, the popular risk of Accurate Prediction mosquito matchmaker disease, kind and the quantity wherein importantly grasping Mosquito Vectors and carry pathogenic micro-organism.That the virus in Mosquito Vectors body is detected to the Main Means of carapuru virus monitoring at present, the method needs a large amount of mosquitoes of collecting, and require have cold chain technology to preserve the integrity of mosquito body inner virus, carry out virus extraction separation, serology and immunological experiment etc. to take back laboratory, or carry out viral RNA analysis [Ritchie SA, et al., 2007].Transport in the wild and carry, the liquid nitrogen container weight that cold chain technology adopts is large, has danger and uses inconvenient; Detecting in analysis, because being easily subject to mosquito self RNA, viral RNA analysis disturbs, and viral separation efficiency is low, and serology and immunology detection sensitivity are not high; Meanwhile, collecting a large amount of mosquitoes, to carry out virus analysis be an effort, expensive work, thereby, need to there is new technology to substitute current carapuru virus Monitoring techniques.
Summary of the invention
The present invention utilizes mosquito to take food counter the telling of saliva in process and discharges viral biological characteristics simultaneously, collects virus, and viral RNA is stored in to carrier more than 7 days at normal temperatures, completes laboratory and detects, and to improve viral separation efficiency, saves man power and material.
Method of the present invention realizes in the following manner, the steps include:
(1) in doubtful mosquito matchmaker Causative virus region, catch 30 ~ 200 one species or different types of mosquito and put into seriatim penicillin bottle, spend the night, after hunger, step to be entered (2);
(2) collect virus by one of following two kinds of methods:
(a) at 24 ~ 27 DEG C of room temperatures, qualitative filter paper is cut into 3cm
2some parts as collecting viral carrier, every part of filter paper central marks drips the position of RNA preservation liquid, get several clean culture dish and add 10% sucrose water, medical cotton ball is immersed in culture dish, filter paper is placed on cotton balls, drip 0.25ml RNA in filter paper mark position again and preserve liquid, by 30 ~ 70 mosquitoes of every part of filter paper, the mosquito of step 1) survival is driven in to the 15 ~ 30cm that is placed with culture dish
3in sarong, allow mosquito take food, change filter paper every day one time, totally 5 ~ 7 days;
Or, (b) step 1) mosquito is collected in penicillin tubule by 48 hours intervals in batches ,-85 DEG C are frozen, detect the infection virus titer of mosquito with plaque test, collect virus liquid; Afterwards, qualitative filter paper is cut into 3cm
2some parts as collecting viral carrier, every part of filter paper central marks drips the position of RNA preservation liquid and virus liquid, get several clean culture dish, place medical cotton ball, filter paper is placed on cotton balls, drip 0.25ml RNA in filter paper mark position and preserve liquid, then get virus liquid and drip 0.25ml in same position;
(3) virus is preserved
Using step 2) filter paper or step 2 that (a) mosquito took food) filter paper that (b) dripped mosquito virus liquid places respectively at 24 ~ 27 DEG C of room temperatures as virus vector, and detected in 7 days;
(4) carrier being taken food by mosquito is extracted to viral RNA
With AxyPrep body fluid viral DNA/RNA small volume of reagent box prescribed manner extraction viral RNA;
(5) pcr amplification and detection
5 ', 3 ' the end conserved sequence primer of the viral bacterium of design PCR reaction system and the PCR reaction conditions of this primer; With PCR reaction product gel imaging and order-checking qualification.
The step 1) of described method is further that the mosquito catching is placed in penicillin bottle and spends the night, and makes its hunger; On next day, drive in the mosquito of survival in sarong, for 8% sucrose water, and 24 ~ 27 DEG C of temperature in sarong, day and night temperature <2 DEG C, relative humidity 70% ~ 80%, light application time 12 ~ 14h, makes its survival.
As aforesaid method, a kind of application is that encephalitis b virus is collected, preserves and detects: step 2) it (b) in the time that the virus titer of the encephalitis b virus liquid infecting is 7. 85 log PFU/ml, collect virus liquid.
Described application, is characterized in that described step 4) pcr amplification PCR reaction system (unit: microlitre) is: MgCl
25 microlitres, Buffer 2.5 microlitres, dNTP 2.5 microlitres, P1 0.5 microlitre, P2 0.5 microlitre, P3 0.5 microlitre, R 1 microlitre, F 1 microlitre, template ribonucleic acid 4 microlitres, H
2o 7.5 microlitres; This PCR reaction system is taking JE-251F and JE-925R as gene amplification primer, wherein:
F: CGT TCT TCA AGT TTA CAg CAT TAG C；
R: CCY RTG TTY CTG CCA AGC ATC CAM CC；
PCR reaction conditions: 1. 50 DEG C 30 minutes; 2. 94 DEG C 5 minutes; 3. 95 DEG C are unwind 30 seconds; 3. anneal 1 minute for 52 DEG C; 4. 72 DEG C are extended 60 seconds; 5. 3. ~ 4. 45 circulation; 6. 72 DEG C are extended 15 minutes; Identified gene amplified fragments is encephalitis b virus band 700bp.
The present invention utilizes mosquito to suck in syrup process when saliva is counter tells and also discharges viral biological characteristics, collects virus, and its viral RNA is stored on carrier, preserves within 7 days and detects for lab analysis at normal temperature.
The present invention has following progress and meaning:
The abduction delivering 1% agarose gel electrophoresis analytical results that number of days is preserved in the stochastic sampling of following Fig. 1 ~ 5 shows: the present invention utilizes mosquito to suck counter the telling of saliva in liquid glucose process and discharges viral biological characteristics simultaneously, the sarong that design and mosquito quantity adapt to and the saturated filter paper of certain density liquid glucose are to collect at normal temperatures virus, and in normal temperature, preserve mosquito viral rna vector and in 7 days, carry out randomly laboratory detection, the trouble of changed the transport of cold chain technology field, carrying and preserving, improve collection virus, preserved efficiency, result is accurate, has saved manpower and materials.
Brief description of the drawings
Fig. 1 is the gel photograph that Culex tritaeniorhynchus of the present invention and Culex quinquefasciatus peroral infection encephalitis b virus normal temperature are preserved the PCR of the 7th day.In figure 1: standard molecular weight protein, 2: the abduction delivering extract of the 7th day.
Fig. 2 is the gel photograph that the present invention reduces Culex tritaeniorhynchus and the 3rd, 4 and 5 days PCR of Culex quinquefasciatus quantity peroral infection encephalitis b virus normal temperature preservation.In figure, 1 is standard molecular weight viral nucleic acid, and 2,3,4 is the abduction delivering extract of the 3rd, 4 and 5 days.
Fig. 3 is that the present invention collects, preserves and detect Culex tritaeniorhynchus by method (b) and Culex quinquefasciatus peroral infection encephalitis b virus normal temperature is preserved latter the 6th day and the gel photograph of 7 days PCR.In figure, 1 is standard molecular weight viral nucleic acid, and 2,3 are respectively the abduction delivering extract of the 6th, 7 days.
Fig. 4 is the gel photograph that PCR that Culex tritaeniorhynchus that the present invention infects encephalitis b virus is preserved at normal temperatures taking filter paper as carrier 1 day detects.In figure, 1 is standard molecular weight viral nucleic acid, and 2 is the abduction delivering extract of the 1st day.
Fig. 5 is that Culex tritaeniorhynchus that the present invention infects encephalitis b virus is preserved respectively the gel photograph that the PCR of 2 days and 7 days detects at normal temperatures taking filter paper as carrier.In figure, 1 is standard molecular weight viral nucleic acid, the extract of 2,3 be respectively abduction delivering the 2nd day and 7 days.
The abduction delivering result that number of days is preserved in Fig. 1 ~ 5 stochastic sampling shows: the object band amplifying with carrier extraction viral RNA is 674bp left and right, conform to the cDNA segment size of the encephalitis b virus primer amplification of estimating, therefore use this carrier and method can complete collection, preservation and the detection to carapuru virus.
Below in conjunction with embodiment, the present invention will be further described, and embodiment comprises but do not limit the content that request of the present invention is protected.
Embodiment
Embodiment 1:
Collect, preserve and detect Culex tritaeniorhynchus and Culex quinquefasciatus peroral infection encephalitis b virus by method (a).
(1) simulate with the mosquito infection experiment of encephalitis street strain virus the mosquito that doubtful mosquito matchmaker Causative virus region catches.
At 30cm
3in sarong, put into the cotton ball of the full suction encephalitis b virus liquid that several grain of rices are large, the encephalitis strain of this virus liquid is the Zhong Shan of encephalitis street strain (Nak) strain, virus titer 7.85 log PFU/ml, to sprout wings latter 1 day, Culex tritaeniorhynchus and 228 of the female mosquitos of Culex quinquefasciatus through 24h hunger drive in sarong, allow mosquito suck (peroral infection), spend the night, next day, the mosquito of survival is driven in penicillin bottle and spent the night, penicillin bottle is placed in 24 ~ 27 DEG C of temperature, day and night temperature <2 DEG C, relative humidity 70% ~ 80% within doors, give illumination 12 ~ 14h, and provide 8% sucrose water, infect 3 days.
Culex using survival is put into penicillin bottle seriatim as the mosquito catching in doubtful mosquito matchmaker Causative virus region, spends the night, and after hunger, step to be entered (2).
(2) collect virus:
At 24 ~ 27 DEG C of room temperatures, qualitative filter paper is cut into 3cm
2viral carrier is collected in totally 3 parts of conducts, every part of filter paper central marks drips the position of RNA preservation liquid, get 3 clean culture dish and add 10% sucrose water, medical cotton ball is immersed in culture dish, filter paper is placed on cotton balls, drip 0.25ml RNA in filter paper mark position again and preserve liquid, by 70 mosquitoes of every part of filter paper, the mosquito of step 1) survival is driven in to the 15 ~ 30cm that is placed with culture dish
3in sarong, allow mosquito take food, change filter paper every day one time, totally 5 days.
(3) virus is preserved:
Using step 2) filter paper that (a) mosquito took food places respectively at 24 ~ 27 DEG C of room temperatures as virus vector, detected at the 7th day.
(4) carrier being taken food by mosquito is extracted to viral RNA:
Use AxyPrep body fluid viral DNA/RNA small volume of reagent box (love pursue progress biotechnology Hangzhou company limited) to comprise: 1. with sterilizing operating scissors, filter paper to be shredded, put into 1.5ml centrifuge tube.Get 500ul MEM liquid and carry out wash-out, vortex oscillation 5 min, centrifugal 5 min of 12,000*g, get supernatant as sample.2. collect sample in 300ul step 1, proceed in 1.5ml centrifuge tube, centrifugal 5 min of 12,000*g, get supernatant as sample.3. add 200ul Buffer V-L, vortex oscillation mixes, and leaves standstill 5 min.4. add 75ul Buffer V-N, vortex oscillation mixes, centrifugal 5 min of 12,000*g.5. supernatant is transferred in new 2ml centrifuge tube, added 300ul Virahol (1% glacial acetic acid), turned upside down 6-8 time, mixes.6. be placed in 2ml centrifuge tube by preparing pipe, the mixed solution immigration of getting in step 4 is prepared in pipe, centrifugal 1 min of 6,000*g.7. abandon filtrate, put and get back in 2ml centrifuge tube preparing pipe, add 500ul Buffer W1A, room temperature leaves standstill 1 minute, centrifugal 1 min of 12,000*g.8. abandon filtrate, put and get back in 2ml centrifuge tube preparing pipe, add 800ul Buffer W2, centrifugal 1 min of 12,000*g.9. put and get back in 2ml centrifuge tube, centrifugal 1 min of 12,000*g preparing pipe.10. be placed in clean 1.5ml centrifuge tube by preparing pipe, add 40-60ul Bufffer TE nuclease-free preparing film central authorities), room temperature leaves standstill 1 min, centrifugal 1 min of 12,000*g, eluted dna/RNA.
(5) pcr amplification
Pcr amplification PCR reaction system (unit: microlitre) comprising: MgCl
25 microlitres, Buffer 2.5 microlitres, dNTP 2.5 microlitres, P1 0.5 microlitre, P2 0.5 microlitre, P3 0.5 microlitre, R 1 microlitre, F 1 microlitre, template ribonucleic acid 4 microlitres, H
2o 7.5 microlitres; This PCR reaction system is taking JE-251F and JE-925R as gene amplification primer, wherein:
F: CGT TCT TCA AGT TTA CAg CAT TAG C；
R: CCY RTG TTY CTG CCA AGC ATC CAM CC。
PCR reaction conditions: 1. 50 DEG C 30 minutes; 2. 94 DEG C 5 minutes; 3. 95 DEG C are unwind 30 seconds; 3. anneal 1 minute for 52 DEG C; 4. 72 DEG C are extended 60 seconds; 5. 3. ~ 4. 45 circulation; 6. 72 DEG C are extended 15 minutes; Identified gene amplified fragments, as Fig. 1, is the about 674bp of encephalitis b virus band.
Embodiment 2:
Reduce mosquito quantity, dwindle sarong to 15cm
3, collect, preserve and detect Culex tritaeniorhynchus and Culex quinquefasciatus peroral infection encephalitis b virus by method (a).
(1) simulate with the mosquito infection experiment of encephalitis street strain virus the mosquito that doubtful mosquito matchmaker Causative virus region catches.
Hunger by 30 female mosquitos of Culex tritaeniorhynchus that sprout wings latter 1 day through 24h, every culex drives in the penicillin bottle of a cotton ball that is placed with the full suction encephalitis b virus liquid that the grain of rice is large, its virus stain is the Zhong Shan of encephalitis street strain (Nak) strain, virus titer is 7.85 log PFU/ml, allow mosquito suck (peroral infection), spend the night.On next day, 24 of the culexs of survival are collected in to 15cm
3in sarong, 24 ~ 27 DEG C of room temperatures, day and night temperature <2 DEG C, relative humidity 70% ~ 80%, cultivates under artificial lighting 12 ~ 14h condition, and 8% sucrose water is provided, and infects 3 days.
Culex using survival is put into penicillin bottle as the mosquito catching in doubtful mosquito matchmaker Causative virus region, spends the night after hunger, and mosquito is driven in to 3cm
2filter paper, is placed on the medical cotton ball of 8% sucrose water soaking, drips 0.25ml RNA and preserves liquid collection virus carrier, allows mosquito take food, and changes carrier every day one time, until infect latter the 11st day.The carrier being taken food by mosquito carries out encephalitis b virus PCR detection.
(2) viral RNA extracts identical with embodiment 1.
(3) pcr amplification and detection are with embodiment 1.
Result: learn from else's experience and infect 24 culexs preservations of encephalitis b virus RNA carrier of 3,4 and 5 days, amplification object band is about 674 bp(Fig. 2).
Embodiment 3:
Collect, preserve and detect Culex tritaeniorhynchus and Culex quinquefasciatus peroral infection encephalitis b virus by method (b).
(1) simulate with the mosquito infection experiment of encephalitis street strain virus the mosquito (identical with embodiment 1) that doubtful mosquito matchmaker Causative virus region catches.
(2) collect virus.
Step 1) is collected in to mosquito-85 in penicillin tubule one by one DEG C frozen, by 48 hours intervals divide 5 batches 211 mosquito plaques tests of totally 13 batch total detect the infection virus titers of mosquitoes, collect virus liquid.Specifically: take out Culex tritaeniorhynchus and the Culex quinquefasciatus of frozen peroral infection, divide into groups by infecting the date, put into mill, with washing 3 times containing 3% dual anti-Hanks liquid, grind, each mill adds 1ml Hanks liquid, make suspension, inoculation has grown up to the BHK21 cell of individual layer.Plaque experiment adopts 6 orifice plates, diameter 35 mm, and every hole inoculation mosquito suspension 0.2 ml, after 37 DEG C of absorption 1h, methylate Mierocrystalline cellulose nutrition coverture covers, CO
2incubator continues to cultivate, and within the 5th day, dyes, calculate plaque number and plaque forming unit with crystal violet solution.Result: have 11 groups to occur virus in 13 211 of batch totals mosquitoes, positive rate is 84.62%, and the virus titer of positive group is higher, and most results are in 4 logarithm left and right (seeing the following form).
Table encephalitis B street strain (strain of middle mountain) infects the empty result of surveying of Culex tritaeniorhynchus
| Detect group number | DAI | Mosquito number of elements | Result (plaque forming unit) |
| 1 | 3 | 15 | 4.23 |
| 2 | 3 | 18 | 3.66 |
| 3 | 5 | 21 | 4.66 |
| 4 | 7 | 20 | 0 |
| 5 | 7 | 19 | 4.81 |
| 6 | 5 | 16 | 3.57 |
| 7 | 5 | 11 | 4.45 |
| 8 | 9 | 17 | 4.63 |
| 9 | 9 | 16 | 3.52 |
| 10 | 11 | 12 | 4.50 |
| 11 | 11 | 14 | 0 |
| 12 | 5 | 16 | 3.71 |
| 13 | 7 | 16 | 3.80 |
Afterwards, qualitative filter paper is cut into 3cm
2viral carrier is collected in totally 3 parts of conducts, every part of filter paper central marks drips the position of RNA preservation liquid and virus liquid, get 3 clean culture dish, a medical cotton ball through 8% sucrose water soaking is placed by each central authorities, filter paper is placed on cotton balls, drip 0.25ml RNA in filter paper mark position and preserve liquid, then get virus liquid and drip 0.25ml in same position.
(3) virus is preserved
Using step 2) filter paper that dripped mosquito virus liquid places respectively at 24 ~ 27 DEG C of room temperatures as virus vector, under 3 parts of filter paper normal temperature, places respectively and detect for 6,7 days.
(4) viral RNA extracts
Identical with embodiment 1 with AxyPrep body fluid viral DNA/RNA small volume of reagent box.
(5) encephalitis b virus pcr amplification and detection
PCR reaction system (total reaction system 25 microlitres) is identical with embodiment 1 with PCR reaction conditions, and gel detection is that normal temperature is preserved the 674bp object band (as Fig. 3) that the filter paper bar of 6 days and 7 days amplifies.
Embodiment 4:
Collect, preserve and detect Culex tritaeniorhynchus peroral infection encephalitis b virus by method (a).
(1) 157 Culex tritaeniorhynchus that catch, infect encephalitis b virus method by embodiment 1 mosquito and infect, and put into seriatim penicillin bottle, spend the night, after mosquito hunger, step to be entered (2).
(2) collect virus by method (a):
At 24 ~ 27 DEG C of room temperatures, qualitative filter paper is cut into 3cm
2viral carrier is collected in totally 3 parts of conducts, every part of filter paper central marks drips the position of RNA preservation liquid, get 3 clean culture dish and add 10% sucrose water, medical cotton ball is immersed in culture dish, filter paper is placed on cotton balls, drip 0.25ml RNA in filter paper mark position again and preserve liquid, driven in by approximately 50 mosquitoes of every part of filter paper the 30cm that is placed with culture dish
3in sarong, allow mosquito take food, change filter paper every day one time, totally 7 days.
(3) virus is preserved
Using step 2) filter paper that took food of mosquito places respectively at 24 ~ 27 DEG C of room temperatures as virus vector, and detected in 1,2 and 7 day.
(4) carrier being taken food by mosquito is extracted to viral RNA identical with embodiment 1.
(5) pcr amplification and detection
PCR reaction system (total reaction system 25 microlitres) is identical with embodiment 1 with PCR reaction conditions.
Fig. 4 is the result of preserving at normal temperatures taking filter paper as carrier 1 day; Fig. 5 is the result of preserving respectively 2 days and 7 days.Experiment shows that the object band amplifying taking carrier extraction viral RNA is as 674bp.
Claims (4)
1. carapuru virus normal temperature Collection and preservation and detection method, comprises the following steps:
(1) in doubtful mosquito matchmaker Causative virus region, catch 30 ~ 200 one species or different types of mosquito and put into seriatim penicillin bottle, spend the night, after hunger, step to be entered (2);
(2) collect virus by one of following two kinds of methods:
(a) at 24 ~ 27 DEG C of room temperatures, qualitative filter paper is cut into 3cm
2some parts as collecting viral carrier, every part of filter paper central marks drips the position of RNA preservation liquid, get several clean culture dish and add 10% sucrose water, medical cotton ball is immersed in culture dish, filter paper is placed on cotton balls, drip 0.25ml RNA in filter paper mark position again and preserve liquid, by 20 ~ 70 mosquitoes of every part of filter paper, the mosquito of step 1) survival is driven in to the 15 ~ 30cm that is placed with culture dish
3in sarong, allow mosquito take food, change filter paper every day one time, totally 5 ~ 7 days;
Or, (b) step 1) is collected in to mosquito-85 in penicillin tubule DEG C frozen, detect the infection virus titer of mosquito with plaque test by 48 hours intervals in batches, collect virus liquid; Afterwards, qualitative filter paper is cut into 3cm
2some parts as collecting viral carrier, every part of filter paper central marks drips the position of RNA preservation liquid and virus liquid, get several clean culture dish, place medical cotton ball, filter paper is placed on cotton balls, drip 0.25ml RNA in filter paper mark position and preserve liquid, then get virus liquid and drip 0.25ml in same position;
(3) virus is preserved
Using step 2) filter paper or step 2 that (a) mosquito took food) filter paper that (b) dripped mosquito virus liquid places respectively at 24 ~ 27 DEG C of room temperatures as virus vector, and detected in 7 days;
(4) carrier being taken food by mosquito is extracted to viral RNA
With AxyPrep body fluid viral DNA/RNA small volume of reagent box prescribed manner extraction viral RNA;
(5) pcr amplification and detection
5 ', 3 ' the end conserved sequence primer of the viral bacterium of design PCR reaction system and the PCR reaction conditions of this primer; With PCR reaction product gel imaging and order-checking qualification.
2. according to the method described in claim 1, it is characterized in that step 1) is further the mosquito catching to be placed in to penicillin bottle spend the night, and makes its hunger; On next day, the mosquito of survival is driven in to step 2) in sarong, 24 ~ 27 DEG C of temperature in sarong, day and night temperature <2 DEG C, relative humidity 70% ~ 80%, light application time 12 ~ 14h, makes its survival.
3. the application of method as claimed in claim 1 or 2 in encephalitis b virus collection, preservation and in detecting, it is characterized in that described step 2) it (b) in the time that the virus titer of encephalitis b virus liquid infecting is 7. 85 log PFU/ml, collect virus liquid.
4. according to the application described in claim 3, it is characterized in that described step 4) pcr amplification PCR reaction system (unit: microlitre) is: MgCl
25 microlitres, Buffer 2.5 microlitres, dNTP 2.5 microlitres, P1 0.5 microlitre, P2 0.5 microlitre, P3 0.5 microlitre, R 1 microlitre, F 1 microlitre, template ribonucleic acid 4 microlitres, H
2o 7.5 microlitres; This PCR reaction system is taking JE-251F and JE-925R as gene amplification primer, wherein:
F: CGT TCT TCA AGT TTA CAg CAT TAG C；
R: CCY RTG TTY CTG CCA AGC ATC CAM CC；
PCR reaction conditions: 1. 50 DEG C 30 minutes; 2. 94 DEG C 5 minutes; 3. 95 DEG C are unwind 30 seconds; 3. anneal 1 minute for 52 DEG C; 4. 72 DEG C are extended 60 seconds; 5. 3. ~ 4. 45 circulation; 6. 72 DEG C are extended 15 minutes; Identified gene amplified fragments is encephalitis b virus band 700bp.
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| CN111088319A (en) * | 2020-03-25 | 2020-05-01 | 深圳市上为生物科技有限公司 | Inactivated virus sample RNA preservation solution and preparation method thereof |
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| ANDREW F.VAN DEN HURK等: "Expectoration of Flaviviruses During Sugar Feeding by Mosquitoes(Diptera:Culicidae)", 《J. MED. ENTOMOL.》, vol. 44, no. 5, 30 September 2007 (2007-09-30), pages 845 - 850 * |
| SONJA HALL-MENDELIN等: "Exploiting mosquito sugar feeding to detect mosquito-borne pathogens", 《PNAS》, vol. 107, no. 25, 22 June 2010 (2010-06-22), pages 11255 - 11259 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111088319A (en) * | 2020-03-25 | 2020-05-01 | 深圳市上为生物科技有限公司 | Inactivated virus sample RNA preservation solution and preparation method thereof |
| CN111088319B (en) * | 2020-03-25 | 2020-07-21 | 深圳市上为生物科技有限公司 | Inactivated virus sample RNA preservation solution and preparation method thereof |
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