CN102608306A - Method for detecting insecticide resistance of schistosoma japonicum eggs to praziquantel in vitro - Google Patents
Method for detecting insecticide resistance of schistosoma japonicum eggs to praziquantel in vitro Download PDFInfo
- Publication number
- CN102608306A CN102608306A CN2011104273856A CN201110427385A CN102608306A CN 102608306 A CN102608306 A CN 102608306A CN 2011104273856 A CN2011104273856 A CN 2011104273856A CN 201110427385 A CN201110427385 A CN 201110427385A CN 102608306 A CN102608306 A CN 102608306A
- Authority
- CN
- China
- Prior art keywords
- praziquantel
- ovum
- drug
- eggs
- schistosoma
- 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
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention relates to a method for detecting insecticide resistance of schistosoma japonicum eggs to praziquantel in vitro, belonging to the technical field of antiparasitic drugs. The method comprises the following steps of: collecting 1,000-2,000 eggs from the feces of schistosoma japonicum infected hosts, respectively immigrating the 1,000-2,000 eggs in a flask of 50mL of normal saline containing praziquantel of 5*10-6mol/L and single control normal saline, and placing the eggs in the dark environment for 24h; then immigrating the eggs in fresh dechlorinated tap water, and hatching miracidia under irradiation of an incandescent bulb of 28 DEG C; after hatching starts, collecting tap water containing the miracidia for one time with a centrifuge tube of 50mL every 30 min; centrifuging after adding few drops of Lugol's iodine solution; collecting the miracidia, observing the miracidia under a microscope, counting, and calculating the hatching rate of eggs; setting normal saline control, and repeating for 3 times; detecting the insecticide resistance of the schistosoma japonicum eggs to praziquantel by observing the hatching rate of eggs. The method is fast, simple, convenient, sensitive and reliable and is suitable for fast detecting and monitoring the insecticide resistance of the schistosoma japonicum to praziquantel on site.
Description
Technical field
A kind of vitro detection schistosoma japonice ovum belongs to the anti-parasite medicine technical field to the drug-fast method of praziquantel.
Background technology
Japanese schistosomiasis is that a kind of harm humans is healthy, influences the serious infectious diseases of socio-economic development.Praziquantel is the current unique commercialization medicine that is used to treat Japanese schistosomiasis.Use repeatedly on a large scale with a kind of pest-resistant medicine for a long time and tend to cause the drug-fast generation of medicine, therefore be necessary to set up field monitoring and detect the drug-fast method of praziquantel.The drug-fast main method of current detection praziquantel is the on-scene care patient in the epidemic-stricken area, assessment chemotherapy result, and this method time is long, patient's succession is low, and exists the under-enumeration leakage to control phenomenon.We have set up a kind of vitro detection schistosoma japonice ovum to the drug-fast technology of praziquantel in the laboratory; Can be easily and fast, the responsive resistance to the action of a drug that detects schistosoma japonice ovum to praziquantel; Solved the technical barrier that the praziquantel resistance to the action of a drug is difficult to detect, helped field monitoring and detect the drug-fast generation of Schistosoma japonicum praziquantel.
Summary of the invention
The purpose of this invention is to provide a kind of vitro detection schistosoma japonice ovum to the drug-fast method of praziquantel, solve field monitoring and detect the drug-fast technical matters of Schistosoma japonicum praziquantel, quick, easy, responsive, reliable.
Technical scheme of the present invention: get two 50 mL flasks, immigration contains 5 * 10 respectively
-6The physiological saline of mol/L praziquantel and single contrast physiological saline, 1000 ~ 2000 worm's ovums will from schistosoma japonicum infection host ight soil, collecting join and contain 5 * 10
-6In the flask of the physiological saline of mol/L praziquantel and single contrast physiological saline, put 24 h in 28 ℃ of dark surrounds, after worm's ovum is moved in the fresh dechlorination tap water; Miracidium is hatched in 28 ℃ of incandescent lamp irradiations down; Per 30 min in hatching beginning back collect the tap water that contains miracidium for 1 time with 50 mL centrifuge tubes, add after the several Lugol's iodine solutions centrifugally, and miracidium is examined under a microscope and counts in collection; Calculate the egg hatch rate; Experiment repetition 3 times through observing the egg hatch rate, detects the resistance to the action of a drug of schistosoma japonice ovum to praziquantel.
Egg hatch rate: the ratio of the miracidium number that miracidium number that the schistosoma japonice ovum of handling through praziquantel hatches and the schistosoma japonice ovum that soaks through physiological saline hatch.
The praziquantel soup is that praziquantel is dissolved in that to be mixed with concentration in 1% dimethyl sulfoxide (DMSO) (DMSO) be 1 * 10
-3Mol/L and water diluted for use.
It is formulated that physiological saline is that 9 g sodium chloride are dissolved in the 1000 mL distilled water.
Lugol's iodine solution is that 5% iodine, 10% potassium iodide and dechlorination water are formulated.
A kind of vitro detection schistosoma japonice ovum adds in 50 mL flasks earlier and contains 5 * 10 the drug-fast method of praziquantel
-6The normal saline solution of mol/L praziquantel, 1000 ~ 2000 worm's ovums will from schistosoma japonicum infection host ight soil, collecting add in the solution, put and place 24 h in 28 ℃ of dark surrounds; After worm's ovum is moved in the fresh dechlorination tap water, the irradiation of 28 ℃ of incandescent lamps is the hatching miracidium down, hatching beginning per 30 min in back contain the tap water of miracidium for 1 time with the collection of 50 mL centrifuge tubes; Add after the several Lugol's iodine solutions centrifugal; Collect miracidium and examine under a microscope and count, calculate the egg hatch rate, the physiological saline contrast is established in experiment; Repeat 6 times, 5 * 10
-6Behind mol/L praziquantel soup effect 24 h, schistosoma japonice ovum Fu Huashuai>0, think that promptly this schistosoma japonice ovum has the resistance to the action of a drug to praziquantel.
Beneficial effect of the present invention: a kind of vitro detection schistosoma japonice ovum is to the drug-fast method of praziquantel; Fast, easy, responsive, reliable; Be fit to field quick detection and the drug-fast generation of monitoring Schistosoma japonicum praziquantel, can solve the technical matters of on-the-spot detection of the Schistosoma japonicum praziquantel resistance to the action of a drug and monitoring.
Embodiment
Embodiment 1: Schistosoma japonicum praziquantel sensitive strain worm's ovum is to the sensitivity testing of praziquantel
Get two 50 mL flasks, immigration contains 5 * 10 respectively
-6The physiological saline of mol/L praziquantel and single with reference to physiological saline; 1000 ~ 2000 worm's ovums to from the sensitive strain infection host ight soil of Schistosoma japonicum Sichuan, collecting join in the flask; Put 24 h in 28 ℃ of dark surrounds; After worm's ovum is moved in the fresh dechlorination tap water, the irradiation of 28 ℃ of incandescent lamps is the hatching miracidiums down.Per 30 min in hatching beginning back collect the tap water that contains miracidium for 1 time with 50 mL centrifuge tubes, add after the several Lugol's iodine solutions centrifugally, and the collection miracidium is examined under a microscope and counts, calculating egg hatch rate.Experiment repetition 3 times.The result shows, at warp 5 * 10
-6Behind mol/L praziquantel effect 24 h, the incubation rate of Sichuan sensitive strain worm's ovum is 0.
Embodiment 2: Schistosoma japonicum praziquantel sensitive strain worm's ovum is to the sensitivity testing of praziquantel
Get two 50 mL flasks, immigration contains 5 * 10 respectively
-6The physiological saline of mol/L praziquantel and single with reference to physiological saline; 1000 ~ 2000 worm's ovums to from the sensitive strain infection host ight soil of Schistosoma japonicum Anhui, collecting join in the flask; Put 24 h in 28 ℃ of dark surrounds; After worm's ovum is moved in the fresh dechlorination tap water, the irradiation of 28 ℃ of incandescent lamps is the hatching miracidiums down.Per 30 min in hatching beginning back collect the tap water that contains miracidium for 1 time with 50 mL centrifuge tubes, add after the several Lugol's iodine solutions centrifugally, and the collection miracidium is examined under a microscope and counts, calculating egg hatch rate.Experiment repetition 3 times.The result shows, at warp 5 * 10
-6Behind mol/L praziquantel effect 24 h, the incubation rate of Anhui sensitive strain worm's ovum is 0.
Embodiment 3: Schistosoma japonicum praziquantel sensitive strain worm's ovum is to the sensitivity testing of praziquantel
Get two 50 mL flasks, immigration contains 5 * 10 respectively
-6The physiological saline of mol/L praziquantel and single with reference to physiological saline; 1000 ~ 2000 worm's ovums to from the sensitive strain infection host ight soil of Schistosoma japonicum Hunan, collecting join in the flask; Put 24 h in 28 ℃ of dark surrounds; After worm's ovum is moved in the fresh dechlorination tap water, the irradiation of 28 ℃ of incandescent lamps is the hatching miracidiums down.Per 30 min in hatching beginning back collect the tap water that contains miracidium for 1 time with 50 mL centrifuge tubes, add after the several Lugol's iodine solutions centrifugally, and the collection miracidium is examined under a microscope and counts, calculating egg hatch rate.Experiment repetition 3 times.The result shows, at warp 5 * 10
-6Behind mol/L praziquantel effect 24 h, the incubation rate of Hunan sensitive strain worm's ovum is 0.
Embodiment 4: Schistosoma japonicum praziquantel sensitive strain worm's ovum is to the sensitivity testing of praziquantel
Get two 50 mL flasks, immigration contains 5 * 10 respectively
-6The physiological saline of mol/L praziquantel and single with reference to physiological saline; 1000 ~ 2000 worm's ovums to from the sensitive strain infection host ight soil of Schistosoma japonicum Jiangsu, collecting join in the flask; Put 24 h in 28 ℃ of dark surrounds; After worm's ovum is moved in the fresh dechlorination tap water, the irradiation of 28 ℃ of incandescent lamps is the hatching miracidiums down.Per 30 min in hatching beginning back collect the tap water that contains miracidium for 1 time with 50 mL centrifuge tubes, add after the several Lugol's iodine solutions centrifugally, and the collection miracidium is examined under a microscope and counts, calculating egg hatch rate.Experiment repetition 3 times.The result shows, at warp 5 * 10
-6Behind mol/L praziquantel effect 24 h, the incubation rate of Jiangsu sensitive strain worm's ovum is 0.
Embodiment 5: Schistosoma japonicum praziquantel resistant strain worm's ovum is to the sensitivity testing of praziquantel
Get two 50 mL flasks, immigration contains 5 * 10 respectively
-6The physiological saline of mol/L praziquantel and single with reference to physiological saline; 1000 ~ 2000 worm's ovums to from the resistant strain infection host ight soil of Schistosoma japonicum Jiangsu, collecting join in the flask; Put 24 h in 28 ℃ of dark surrounds; After worm's ovum is moved in the fresh dechlorination tap water, the irradiation of 28 ℃ of incandescent lamps is the hatching miracidiums down.Per 30 min in hatching beginning back collect the tap water that contains miracidium for 1 time with 50 mL centrifuge tubes, add after the several Lugol's iodine solutions centrifugally, and the collection miracidium is examined under a microscope and counts, calculating egg hatch rate.Experiment repetition 3 times.The result shows, at warp 5 * 10
-6Behind mol/L praziquantel effect 24 h, the incubation rate of Jiangsu resistant strain worm's ovum is (11.2 ± 3.5) %.
Embodiment 6: Schistosoma japonicum praziquantel resistant strain worm's ovum is to the sensitivity testing of praziquantel
Get two 50 mL flasks, immigration contains 5 * 10 respectively
-6The physiological saline of mol/L praziquantel and single with reference to physiological saline; 1000 ~ 2000 worm's ovums to from the resistant strain infection host ight soil of Schistosoma japonicum Hunan, collecting join in the flask; Put 24 h in 28 ℃ of dark surrounds; After worm's ovum is moved in the fresh dechlorination tap water, the irradiation of 28 ℃ of incandescent lamps is the hatching miracidiums down.Per 30 min in hatching beginning back collect the tap water that contains miracidium for 1 time with 50 mL centrifuge tubes, add after the several Lugol's iodine solutions centrifugally, and the collection miracidium is examined under a microscope and counts, calculating egg hatch rate.Experiment repetition 3 times.The result shows, at warp 5 * 10
-6Behind mol/L praziquantel effect 24 h, the incubation rate of Hunan resistant strain worm's ovum is (17.5 ± 6.7) %.
Claims (6)
1. a vitro detection schistosoma japonice ovum is characterized in that getting two 50 mL flasks to the drug-fast method of praziquantel, moves into respectively to contain 5 * 10
-6The physiological saline of mol/L praziquantel and single contrast physiological saline, 1000 ~ 2000 worm's ovums will from schistosoma japonicum infection host ight soil, collecting move into respectively and contain 5 * 10
-6In the flask of the physiological saline of mol/L praziquantel and single contrast physiological saline; Put 24 h in 28 ℃ of dark surrounds, after worm's ovum is moved in the fresh dechlorination tap water, the irradiation of 28 ℃ of incandescent lamps is the hatching miracidiums down; Hatching beginning per 30 min in back collect the tap water that contains miracidium for 1 time with 50 mL centrifuge tubes; Add after the several Lugol's iodine solutions centrifugally, collect miracidium and examine under a microscope and count, calculate the egg hatch rate; If the physiological saline contrast repeats 3 times,, detect the resistance to the action of a drug of schistosoma japonice ovum to praziquantel through observing the egg hatch rate.
2. ask 1 described vitro detection schistosoma japonice ovum to the drug-fast method of praziquantel according to right power; It is characterized in that the egg hatch rate, the ratio of the miracidium number that miracidium number that the schistosoma japonice ovum of handling through praziquantel hatches and the schistosoma japonice ovum that soaks through physiological saline hatch.
3. ask 1 described vitro detection schistosoma japonice ovum to the drug-fast method of praziquantel according to right power, it is characterized in that the praziquantel soup is that praziquantel is dissolved in that to be mixed with concentration among the 1% dimethyl sulfoxide (DMSO) DMSO be 1 * 10
-3Mol/L and water diluted for use.
4. ask 1 described vitro detection schistosoma japonice ovum to the drug-fast method of praziquantel according to right power, it is formulated to it is characterized in that physiological saline is that 9 g sodium chloride are dissolved in the 1000 mL distilled water.
5. ask 1 described vitro detection schistosoma japonice ovum to the drug-fast method of praziquantel according to right power, it is characterized in that Lugol's iodine solution is that 5% iodine, 10% potassium iodide and dechlorination water are formulated.
Like claim 1,2,3 or 4 described vitro detection schistosoma japonice ovums to the drug-fast method of praziquantel, it is characterized in that in 50 mL flasks, adding earlier containing 5 * 10
-6The normal saline solution of mol/L praziquantel, 1000 ~ 2000 worm's ovums will from schistosoma japonicum infection host ight soil, collecting add in the solution, put and place 24 h in 28 ℃ of dark surrounds; After worm's ovum is moved in the fresh dechlorination tap water, the irradiation of 28 ℃ of incandescent lamps is the hatching miracidium down, hatching beginning per 30 min in back contain the tap water of miracidium for 1 time with the collection of 50 mL centrifuge tubes; Add after the several Lugol's iodine solutions centrifugal; Collect miracidium and examine under a microscope and count, calculate the egg hatch rate, the physiological saline contrast is established in experiment; Repeat 6 times, 5 * 10
-6Behind mol/L praziquantel soup effect 24 h, schistosoma japonice ovum Fu Huashuai>0, think that promptly this schistosoma japonice ovum has the resistance to the action of a drug to praziquantel.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011104273856A CN102608306A (en) | 2011-12-20 | 2011-12-20 | Method for detecting insecticide resistance of schistosoma japonicum eggs to praziquantel in vitro |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011104273856A CN102608306A (en) | 2011-12-20 | 2011-12-20 | Method for detecting insecticide resistance of schistosoma japonicum eggs to praziquantel in vitro |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN102608306A true CN102608306A (en) | 2012-07-25 |
Family
ID=46525846
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2011104273856A Pending CN102608306A (en) | 2011-12-20 | 2011-12-20 | Method for detecting insecticide resistance of schistosoma japonicum eggs to praziquantel in vitro |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102608306A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105385686A (en) * | 2015-12-15 | 2016-03-09 | 中国人民解放军军事医学科学院基础医学研究所 | Improved LAMP method-based schistosoma japonicum katsurada nucleic acid detection kit and method |
| CN106172263A (en) * | 2016-08-31 | 2016-12-07 | 江苏省血吸虫病防治研究所 | A kind of semi-automatic human excrement schistosoma japonice ovum hatching apparatus and hatching method thereof |
-
2011
- 2011-12-20 CN CN2011104273856A patent/CN102608306A/en active Pending
Non-Patent Citations (3)
| Title |
|---|
| 李伟等: "《日本血吸虫虫卵杀灭剂筛选及快速杀灭技术的研究 I化学杀卵剂的筛选》", 《中国血吸虫病防治杂志》 * |
| 梁幼生等: "《血吸虫对吡喹酮抗药性的研究 II曼氏血吸虫吡喹酮敏感株与抗性株虫卵和毛蚴对吡喹酮的反应性》", 《中国血吸虫病防治杂志》 * |
| 梁幼生等: "《血吸虫对吡喹酮抗药性的研究XI日本血吸虫中国大陆现场分离株虫卵 毛蚴 尾蚴对吡喹酮的敏感性》", 《中国血吸虫病防治杂志》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105385686A (en) * | 2015-12-15 | 2016-03-09 | 中国人民解放军军事医学科学院基础医学研究所 | Improved LAMP method-based schistosoma japonicum katsurada nucleic acid detection kit and method |
| CN105385686B (en) * | 2015-12-15 | 2019-03-22 | 中国人民解放军军事医学科学院基础医学研究所 | A kind of Schistosoma japonicum kit for detecting nucleic acid and detection method based on improvement LAMP method |
| CN106172263A (en) * | 2016-08-31 | 2016-12-07 | 江苏省血吸虫病防治研究所 | A kind of semi-automatic human excrement schistosoma japonice ovum hatching apparatus and hatching method thereof |
| CN106172263B (en) * | 2016-08-31 | 2021-12-21 | 江苏省血吸虫病防治研究所 | Semi-automatic human-excrement schistosoma japonicum egg hatching device and hatching method thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Kazer et al. | Evolution and diversity of immune responses during acute HIV infection | |
| CN202289094U (en) | Transfusion alarm | |
| Patil et al. | Prevalence of leptospirosis among dogs and rodents and their possible role in human leptospirosis from Mumbai, India | |
| Ma et al. | Decline of SARS-CoV-2-specific IgG, IgM and IgA in convalescent COVID-19 patients within 100 days after hospital discharge | |
| CN102608306A (en) | Method for detecting insecticide resistance of schistosoma japonicum eggs to praziquantel in vitro | |
| CN102329879B (en) | Rapid identification method of Brucellosis aerosol by using fluorescent quantitative PCR (polymerase chain reaction) | |
| CN102936600A (en) | A549 nude mouse model of stably expressed luciferase and building and application thereof | |
| CN109269997A (en) | Distributed Faecal occult blood detection device and its method | |
| Praharaj et al. | Enteropathogens and gut inflammation in asymptomatic infants and children in different environments in southern India | |
| CN104928288A (en) | Nested primer used for early warning of enterocytozoon hepatopenaei of Chinese cultured shrimps and application of nested primer | |
| CN102323404A (en) | Autoimmunity antibody detection kit and detection method | |
| Sachdeva et al. | Preparing for mpox resurgence: surveillance lessons from outbreaks in Toronto, Canada | |
| Steward | Anthelmintic studies: I. A controlled critical entero-nemacidal test | |
| Tabrizi et al. | Determination of triamterene in human plasma and urine after its cloud point extraction | |
| Pereira et al. | Pharmacokinetic and pharmacodynamic evaluation following vaginal application of IQB3002, a dual-chamber microbicide gel containing the nonnucleoside reverse transcriptase inhibitor IQP-0528 in rhesus macaques | |
| CN102876811A (en) | RT-LAMP (reverse transcription loop-mediated isothermal amplification) detection kit and RT-LAMP detection method for SVCV (spring viremia of carp virus) | |
| CN109187964A (en) | A kind of method and its application purifying detection simultaneously to avian leukosis and salmonellosis of chicken using one piece of hatching egg | |
| CN103728458A (en) | Applications of rabbit encephalitozoon cuniculi spore wall protein SWP1 to preparation of reagent for diagnosing or detecting rabbit encephalitozoon cuniculi infection | |
| Gomes et al. | Accuracy of the 14C-urea breath test for the diagnosis of Helicobacter pylori | |
| Pu et al. | Serologic evidence of prevalent avian H3 subtype influenza virus infection in chickens | |
| CN109295200A (en) | The Biological Detection marker of High Linear energy transmission ray radiation and its application | |
| CN102608305A (en) | Method for detecting insecticide resistance of schistosoma japonicum miracidia to praziquantel in vitro | |
| Miura et al. | Evaluation of quantitative polymerase chain reaction for the detection of Toxoplasma gondii oocysts shed by cats | |
| Sun et al. | Cheng's Juanbi Decoction enhances autophagy in rheumatoid arthritis fibroblast-like syn-oviocytes by suppressing the PI3K/Akt/mTOR signal axis | |
| CN102520149A (en) | Method for detecting medicine resistance of schistosoma japonicum cercariae to praziquantel in vitro |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20120725 |