GB 15977-1995 Diagnostic criteria and management principles for schistosomiasis
Some standard content:
National Standard of the People's Republic of China
Diagnostic criteria and principles ofmanagement of schistosomiasis
Diagnostic criteria and principles ofmanagement of schistosomiasisGB 15977-1995
This standard is formulated in accordance with the Law of the People's Republic of China on the Prevention and Control of Infectious Diseases and the Measures for the Implementation of the Law of the People's Republic of China on the Prevention and Control of Infectious Diseases. Subject content and scope of application
This standard specifies the diagnostic criteria, treatment methods and prevention and control principles for schistosomiasis at various stages. This standard is applicable to the prevention and control of schistosomiasis by professional institutions in epidemic areas and the diagnosis and treatment of schistosomiasis patients by medical and health institutions of all levels and types across the country.
2 Diagnostic principles
Diagnosis is made based on the history of contact with infected water, combined with the main symptoms and signs such as fever, diarrhea, hepatomegaly, liver fibrosis and portal hypertension, as well as parasitological examinations, serum immunological examinations, and blood test results. 3 Diagnostic criteria
3.1 Acute schistosomiasis
3.1.1 History of contact with infected water 2 weeks to 3 months before onset. 3.1.2 Fever, hepatomegaly and peripheral blood eosinophilia are the main features, accompanied by tenderness in the liver area, splenomegaly, cough, abdominal distension and diarrhea.
3.1.3 Stool examination reveals schistosome eggs or schistosomes (see Appendix A for details). 3.1.4 Positive serum immunoreactions such as ring egg, hemagglutination, enzyme labeling, and latex (ring egg precipitation test ring sedimentation rate ≥3% and (or) indirect hemagglutination titer ≥1:10. Enzyme labeling reaction is positive, and latex agglutination test titer ≥1:10) (see Appendix B for details). Suspected cases: meet 3.1.1 and 3.1.2. Confirmed cases: Suspected cases add 3.1.3.
Clinical diagnosis: Suspected cases add 3.1.4.
3.2 Chronic schistosomiasis
3.2.1 Living in or having been to an endemic area with a history of contact with infected water. 3.2.2 Asymptomatic, or with occasional abdominal pain, diarrhea, or bloody stools. Most cases are accompanied by an enlarged liver, mainly in the left lobe, and a few are accompanied by an enlarged spleen. 3.2.3 Schistosome eggs or hairy spiders are found in stool examination, or schistosome eggs are found in rectal biopsy in patients without a history of treatment, and live eggs or recently degenerated eggs are found in patients with a history of treatment (see Appendix A for details).
3.2.4 Patients with no history of schistosomiasis treatment or who have been treated for more than 3 years, with a ring ovum sedimentation test ring sedimentation rate ≥3% and (or) indirect hemagglutination test titer ≥1:10, positive enzyme-labeled reaction, and latex agglutination test titer ≥1:10; patients who have not been treated or have been treated for more than 1 year have positive serum schistosomiasis circulating antigens (see Appendix B for details).
Suspected cases: meet 3.2.1 and 3.2.2. Confirmed cases: add 3.2.3 to suspected cases.
Clinical diagnosis: add 3.2.4 to suspected cases. Approved by the State Administration of Technical Supervision on December 15, 1995 and implemented on July 1, 1996
3.3 Late Schistosomiasis
GB159771995
3.3.1 History of long-term or repeated contact with infected water, or clear history of schistosomiasis treatment. 3.3.2 Clinical symptoms and signs of portal hypertension, or dwarfism or colonic granuloma. 3.3.3 Eggs or hairy spiders are found in fecal examination, or schistosomiasis eggs are found in rectal biopsy in patients without treatment history, and live eggs or recently degenerated eggs are found in patients with treatment history (see Appendix A for details).
3.3.4 Serological diagnosis is positive, see 3.2.4 for standards. Suspected cases: meet 3.3.1 and 3.3.2. Confirmed cases: add 3.3.3 for suspected cases.
Clinical diagnosis: add 3.3.4 for suspected cases.
4 Treatment
4.1 Chemotherapy
4.1.1 Individual chemotherapy
See Appendix C for details.
4.1.1.1 Acute schistosomiasis - generally use praziquantel at a total dose of 120mg/kg (140mg/kg for children) for 6 days. 4.1.1.2 Chronic schistosomiasis generally uses praziquantel 40mg/kg once or twice a day. 4.1.1.3 Late schistosomiasis generally uses praziquantel at a total dose of 60mg/kg, divided into 3~~6 doses within 1 to 2 days. 4.1.2 Group chemotherapy
4.1.2.1 Through fecal sampling survey, in administrative villages, in areas where the schistosomiasis infection rate is higher than 20%, no pathogen or serological examination is conducted, and general treatment is conducted for people aged 5 years and above and under 60 years old who have a history of contact with infected water in the year and have no contraindications. General treatment uses 40mg/kg of praziquantel taken at once or in two doses.
4.1.2.2 For epidemic areas where the infection rate in the sampling survey is below 20%, fecal examination or serum immune response examination should be conducted. Those who meet one of the conditions in 3.2.3 or 3.2.4 should be treated with 40mg/kg of praziquantel. 4.1.2.3 Fishermen, boatmen, herdsmen and other people who frequently contact infected water should not be examined, and treatment should be conducted once or twice a year at the above dosage. 4.1.2.4 Livestock (mainly cattle and buffaloes) in moderate and severe epidemic areas should be chemotherapy once a year, which should be carried out simultaneously with chemotherapy for the population. Livestock chemotherapy: Cattle can take 30mg/kg of veterinary praziquantel once a day, buffaloes can take 25mg/kg once a day, and pigs can take 30mg/kg once a day. 4.1.2.5 Chemotherapy time: In endemic areas, human chemotherapy can generally be carried out after the end of the infection season in winter. Livestock chemotherapy is best carried out simultaneously with human chemotherapy.
4.2 Snail control
4.2.1 For endemic areas with large snail areas, it is generally advisable to treat snail habitats in susceptible areas, mainly using the chemical snail control drug chloranilamide to kill snails, 1 to 2 times a year. It is supplemented by necessary snail control methods that change the living environment of snails as much as possible in combination with production. 4.2.2 For endemic areas with limited snail areas, the main method is to change the environment in combination with production and water conservancy, supplemented by drug killing. 4.3 Protection
In places with high density of infectious snails, especially in rivers, lakes, islands and beaches, schistosomiasis protection must be done well. Protection methods should be determined according to local conditions. This includes using snail killers to kill snails in the water, changing farming methods, or conducting protective snail killing, or using skin anti-snail agents to avoid or reduce people's contact with infected water, kill or remove snails in the water, or prevent snail invasion. If you have already come into contact with infected water, you can get early treatment.
4.4 Safe water use
Protect drinking water sources, reduce or avoid human and animal feces pollution; dig wells or build rural tap water; or use heating or drugs to kill snails before use to reduce or prevent snail invasion. 4.5 Feces management
On the basis of finding out how local feces pollute water sources, combined with rural health construction, formulate specific measures for the management and treatment of feces to prevent feces from polluting water sources and kill schistosome eggs in feces. 236
A1 Stool examination
A1.1 Nylon bag egg collection and hatching method
GB 15977—1995
Appendix A
Pathological examination
(Supplement)
Operation steps: Take about 30g of the subject's stool and put it in a copper wire sieve of 40~60 mesh/in. The copper wire sieve is placed on the mouth of a nylon silk bag (260 mesh/in) with an iron clip at the bottom. Water is poured to adjust the slurry so that the fecal liquid is directly filtered into the nylon silk bag. Then remove the copper wire sieve, continue to pour water to rinse the fecal residue in the bag, and use bamboo chopsticks to gently scrape the outside of the bag to help filtration until the filtrate becomes clear. Remove the iron clip at the bottom of the bag and rinse the sediment in the bag into a conical flask. If additional precipitation microscopic examination is required, 34 drops of sediment can be taken from the flask and placed on a glass slide, and then smeared into a smear. The smear surface should occupy 2/3 of the area of the glass slide. The thickness of the smear should be based on the standard that the printed font can be clearly seen through the smear, and the smear should be placed under a low-power microscope for examination. The time for full-film microscopic examination should not be less than 2 minutes, and at least two smears should be examined for each stool. Schistosoma eggs and other worm eggs should be carefully identified during microscopic examination. Then add water to the triangular flask containing fecal sediment to 1 cm from the bottle mouth, put it in the incubation room (box) or incubate it at room temperature. After a certain period of time, take out the flask and observe the hair. Generally, it is necessary to observe 2 to 3 times, and the observation time varies with the temperature. When the temperature is high, the hair will hatch earlier; when the temperature is low, the hair will hatch later. When the temperature is over 30℃, the first observation can be carried out after 0.5-1h. If the result is negative, the second observation can be carried out after 4h, and the third observation can be carried out after 8h. If the result is negative after 3 times, it is judged as negative. When the temperature is between 26 and 30℃, the observation can be started after 4h after incubation, and the result can be observed again after 8h and 12h if the result is negative. When the temperature is between 20 and 25℃, the first observation can be carried out after 8h, and the second observation can be carried out after 12h. If the incubation is carried out at natural temperature, the temperature difference between day and night is large, and the observation can be carried out again in the morning after the operation. When the temperature is 20℃, the observation can be carried out after 12h and 24h respectively. Generally, when the room temperature is around 20℃, the incubation can be carried out at natural temperature without heating. When observing hair, the flask should be facing the light source and lined with black cardboard. Pay attention to the difference between hair and protozoa in water. If in doubt, it can be sucked out with a capillary pipette and identified under a microscope. A1.2 Modified Kato-Katz thick smear method
A1.2.1 Operation steps
A1.2.1.1. Place a nylon silk sheet on the fecal sample to be tested, and use a soft plastic scraper to gently scrape the nylon sheet. The fecal residue will be exposed from the micropores of the silk sheet to the surface of the silk sheet.
A1.2.1.2 Place a quantitative plate (3cm×4cm×2.5cm, the diameter of the circular hole in the plate is 3.5mm, and after scraping, the hole can hold 41.1mg of feces) in the middle of the slide, and use the scraper to scrape the fine feces residue from the nylon silk sheet and fill it into the central hole of the quantitative plate. Fill it up and scrape it flat. A1.2.1.3 Carefully lift the quantitative plate, and the fecal sample will remain on the slide. A1.2.1.4 Take a hydrophilic glass paper (30mm×30mm) that has been soaked in glycerol-malachite green solution for 24 hours, cover it on the feces, and use a rubber stopper or another glass slide to cover the glass paper and gently press it to make the feces evenly spread to the edge of the glass paper. A1.2.1.5 After numbering, place overnight at room temperature of 25℃ and relative humidity of 75%, and examine under a microscope. The transparent time for thin-shelled eggs, such as hookworm eggs, is preferably 0.5 to 1.0 hours, and the longest time should not exceed 2 hours. Otherwise, they will be missed due to excessive transparency. A1.2.1.6 At least 2 smears should be made for each fecal sample. The average number of eggs detected in each piece under microscopic examination multiplied by 24 is the number of eggs in 1g of feces (EPG).
A1.3 Egg collection transparency method
Operation steps: After fully stirring the feces, take 5g and place it in a pond porcelain cup, add water to make fecal liquid. Filter the fecal liquid through a 60 mesh/in copper wire sieve and pour water into two stacked nylon bags (bag depth 20cm, bag opening diameter 8cm, outer bag 260 mesh/in, inner bag 120 mesh/in). Then remove the copper wire sieve, continue to rinse the fecal residue in the bag with water, and gently shake the bag to accelerate filtration until the filtrate becomes clear. Use a medicine spoon to scrape all the sediment in the outer bag and divide it into smears. On the sediment smear, cover the hydrophilic glass paper (2cm×5cm) that has been soaked in glycerol-malachite green solution for 24 hours, press the spoon with a toothpick, leave it at room temperature overnight, and examine it under a microscope the next day. The number of eggs obtained from all the sediments is the same, and then divided by 5 to obtain the number of eggs per gram of feces (EPG).
A2 Rectal biopsy
GB 15977---1995
Performed according to hospital routine. This method can be used for the diagnosis of suspected patients in the hospital, but should not be used for general screening. A3 Pathological examination of liver and other tissue biopsy or surgical specimens For those with no medical history, pathological examination of liver and other tissue biopsy or surgical specimens can confirm the diagnosis of schistosoma eggs. Appendix B
Serum immunological examination
(Supplement)
B1 Ring egg precipitation test (COPT)
B1.1 Common methods
B1.1.1 Eggs: Heat-treated ultrasonic dried egg powder. The ring precipitation rate is >30% when tested with heavily infected rabbit serum (rabbit serum inoculated with 1500-2000 tail butterflies, 42 days old).
B1.1.2 Operation method: First, use melted paraffin to draw two wax lines 20mm apart at both ends of a clean glass slide, add 2 drops of serum (0.05-0.10mL) of the subject between the wax lines, then pick about 100-150 dried eggs with a needle, add them to the serum, mix well, cover with a 24mm×24mm cover glass, seal with paraffin on all sides, and place in a 37℃ incubator. After 48-72 hours, observe the reaction results with a low-power (80-100×) microscope. Suspected ones should be identified under a high-power (400×) microscope. To simplify the operation, you can also use prefabricated dried egg PVC membranes. Just add serum, keep it in a humidified box at 37℃ for 24 hours, take it out, pour off the serum, add a small amount of saline and observe the reaction under a microscope. B1.1.3 Reaction standard: A typical positive reaction is a bubble-like, finger-like or slender, curly ribbon-like precipitate with neat edges and obvious refraction. The bubble-like sediment must be larger than 10um (equivalent to the size of two red blood cells) to be considered positive. For positive specimens, 100 mature eggs should be observed and their sedimentation rate calculated; for negative specimens, the entire specimen must be viewed. Negative reaction: The area around the eggs is smooth and has no sediment; or there are bubble-like sediments smaller than 10μm. The intensity and ring sedimentation rate of positive reactions:
a. "Ten" The area of bubble-like and finger-like sediments around the eggs is less than 1/4 of the area of the eggs; the slender and curly ribbon-like sediments are smaller than the long diameter of the eggs.
b. "Ten-ten" The area of bubble-like and finger-like sediments around the eggs is greater than 1/4 of the area of the eggs; the slender and curly ribbon-like sediments are equal to or exceed the long diameter of the eggs.
c. "Ten-ten+" The area of bubble-like and finger-like sediments around the eggs is greater than 1/2 of the area of the eggs; the slender and curly ribbon-like sediments are equal to or exceed twice the long diameter of the eggs.
Number of positive eggs
Ring sedimentation rate (%) = Number of eggs observed on the slide × 100B1.2 Double-sided adhesive tape method
(B1)
B1.2.7 Eggs: Heat-treated ultrasonic dried egg powder. Using 25mg/mL. of heavily infected rabbit plaque Y-globulin as standard serum, the ring sedimentation rate > 30% is qualified.
B1.2.2 Operation method: Take a domestic double-sided adhesive tape with double circular holes (thickness of about 150μm, circular hole diameter 16mm, spacing 8mm), first peel off the cover paper on one side, and then stick it on the slide. The adhesive tape must be tightly adhered to the slide without leaving any gaps. Then remove the cover paper on the adhesive strip, use a suitable dropper to drop 2 drops of about 50μl serum into the round hole, cover with a cover glass (22mm×22mm), place in a 37℃ incubator, and observe the results for 48-72 hours. B1.2.3 Reaction standard: Same as B1.1.3.
B2 Indirect hemagglutination test (IHA)
B2.1 Common methods
GB159771995
B2.1.1 Antigen: Sheep red blood cells sensitized with soluble schistosome egg antigens preliminarily purified with dextran gel G100. The sheep red blood cells used are first sensitized with 2.5% glutaraldehyde and 1:5000 acid solution before sensitization. The sensitized red blood cells are prepared with a 5% suspension in pH7.2 PBS containing 10% sucrose and 1% normal rabbit serum, and then packaged and sealed at low pressure freeze-dried. Each batch of sensitized red blood cells is tested for titer, and the titer is qualified when it reaches 1:1280-2560.
B2.1.2 Operation method:
B2.1.2.1 Open the safe, dilute and mix each tube with 1mL of distilled water for later use. B2.1.2.2 Use a micropipette to add 4 drops (0.025mL/drop) of normal saline to the second well of the first row of the U-shaped micro-hemagglutination reaction plate, the third well is blank, and 1 drop is added to the fourth well.
B2.1.2.3 The serum to be tested is stored in the first well, and 1 drop of serum is drawn from it and added to the second well. After fully mixing, two drops are drawn and 1 drop is added to the third and fourth wells. After mixing in the fourth well, 1 drop is discarded so that the serum dilution ratios of the third and fourth wells are 1:5 and 1:10 respectively. B2.1.2.4 Use a quantitative pipette to absorb the sensitized red blood cell suspension, add 1 drop to each of the third and fourth wells, immediately rotate and shake for 2 minutes, leave it at room temperature for about 1 hour, and observe the results.
B2.1.2.5 Each test should have positive serum and saline as positive and negative controls. B2.1.3 Result judgment:
B2.1.3.1 Negative reaction is that all red blood cells sink to the bottom of the well, and a small, dense dot with smooth edges can be seen by the naked eye. B2.1.3.2 Positive reaction.
++++Red blood cells form a thin layer of agglutination, with irregular wrinkles on the edges. Ten Ten Ten Red blood cells form a thin layer of agglutination, filling the entire bottom of the well. ++
Red blood cells form a thin layer of agglutination, with a smaller area than the ten Ten Ten. Most of the red blood cells sink to the bottom of the well to form a dot, surrounded by a small number of agglutinated red blood cells, and the surrounding area is blurred to the naked eye (or a more obvious blank spot appears in the middle).
B2.1.4 Reaction standard: A positive reaction with serum diluted 1:10 can be judged as a schistosomiasis patient. B2.2 Diaminophenol sensitization method
B2.2.1 Antigen: It is a mixture of soluble egg antigen (SEA) of Schistosoma japonicum and urea soluble antigen (AUA) of adult worms, with a ratio of SEA10mg/L + AUA20mg/L. The sheep or human type \()\ red blood cells used are sensitized with 5% glutaraldehyde and sensitized with diaminophenol. PBS containing 10% sucrose and 1% normal rabbit serum at pH7.2 is added to the sensitized live red blood cells, and the cells are packaged and freeze-dried for storage. Each batch of sensitized red blood cells is titered with positive reference serum and the titer reaches 1:640~1280. Those with no agglutination in negative serum and saline control are qualified. B2.2.2 Operation method
B2.2.2.1 Each tube of freeze-dried sensitized red blood cells is 0.5mL. Dilute with 1mL of normal saline before use. B2.2.2.2 In a \V”-shaped (90°) blood coagulation plate, dilute the test serum with normal saline at a ratio of 1:5 to 1:640. B2.2.2.3 Except for the 1:5 dilution well, add drops (0.03mL) sensitized red blood cells, shake for 30s, and observe the reaction results for 60-90min.
B2.2.3 Reaction standard: A positive reaction is when the wells show strong agglutination (+++) at a dilution of 1:10. B3 Enzyme-linked immunosorbent assay (ELISA)
B3.1 Common methods
B3.1.1 Antigen: Soluble egg antigen. B3.1.2 Operation method
B3.1.2.1 Add 0.2mL of egg antigen diluted 1:3000 (or 1:1000, depending on the antigen titer) with pH 9.6 carbonate buffer to the concave wells of a micro-polystyrene plastic plate and place at 4°C overnight. 230
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B3.1.2.2 Pour off the antigens the next day, wash three times with phosphate buffered saline (PBS/T pH 7.4, 0.01 mol/L) containing 0.05% Tween-20, 5 min each time,
B3.1.2.3 Add 0.2 mL of the subject's serum diluted 1:200 with PBS/T to the concave wells, and incubate at 37°C for 2 h. B3.1.2.4 Pour off the serum, wash three times with PBS/T, 5 min each time. B3.1.2.5 Add 0.2 mL of horseradish peroxidase (HRP)-labeled conjugate diluted 1:1000 with PBS/T, and incubate at 37°C for 2 h. B3.1.2.6 Pour off the enzyme-labeled conjugate, wash three times with PBS/T, 5 min each time. B3.1.2.7 Add 0.2mL o-phenylenediamine (OPD) substrate solution C10mgOPD+pH5.0 citric acid buffer 25mL+30% hydrogen peroxide H2O,) 10μLJ0.2mL, 37℃, 30min. B3.7.2.8 Add 0.05mL of 2mol/L sulfuric acid (HzSO) to each well to stop the reaction. B3.1.2.9 Read the optical density (OD) value at 492nm on the enzyme-labeled colorimeter. B3.1.2.10 Set a positive reference serum control for each experiment, and calculate its OD value in the same way. Use this to calibrate the sample serum OD value. Sample serum OD value
OD correction value - reference concave serum OD value
B3.1.3 Reaction standard: According to the results of the Dynatech enzyme-labeled colorimeter, an OD value ≥0.5 is positive for Schistosoma japonicum. B3.2 ELISA using purified antigens
B3.2.1 Antigen: Soluble egg antigen (AEA) of Schistosoma japonicum purified by 50% to 75% saturated ammonium sulfate. B3.2.2 Operation method:
B3.2.2.1 Wash the 40-well polystyrene plastic plate coated with AEA antigen once with PBS/T (pH 7.4) before use. · (B2)
B3.2.2.2 Dilute the serum with PBS/T to 1:200, add 0.1 mL to each well, add 2 wells for each serum, place in a covered humidified box, and incubate at 20-37°C for 30 minutes.
B3.2.2.3 Remove all the liquid in the wells of the reaction plate, fill each well with PBS/T, remove all the liquid in the wells after 3-5 minutes, and repeat this washing 3 times. bzxz.net
B3.2.2.4Add 0.1mL of 1:1000 diluted horseradish peroxidase (HRP) enzyme conjugate to each well, incubate and wash as in B3.2.2.2~B3.2.2.3.
B3.2.2.5Add 0.1mL of OPD substrate solution (OPD 20mg + PH5.0 citric acid-phosphate buffer 50mL, add 30% H,O, 20μL before use), incubate as in B3.2.2.2. B3.2.2.6Add 2mol/L H,SO, 25μL to each well to terminate the reaction. B3.2.2.7Read the extinction value at 492nm with an enzyme-labeled detector. Calculate the average OD value of the two wells for each sample, and then calibrate it with standard serum.
OD value of the sample to be tested = OD value of standard serum × measured OD value of the sample to be tested. (B3)
B3.2.3 Reaction standard: The OD value of each plate of standard serum should be controlled within the range of 0.79 to 1.15. If it fails to reach this quality control range, it should be retested.
Specimen OD value ≥ 0.5 is considered positive.
B3.3PVC film rapid enzyme-linked immunosorbent assay (ELISA) B3.3.1 Antigen: Soluble egg antigen (SEA) of Schistosoma japonicum and soluble egg antigen (JEU) of 8 mol/L urea are mixed in equal amounts. B3.3.2 Operation method:
B3.3.2.1 Before the experiment, number the back of the PVC film coated with antigen, wash once with washing solution (0.05% Tween saline), and then add 0.2mL of PBS/T to each well
B3.3.2.2 Add the test serum and reference serum according to the number (one negative control and one positive control for each batch), 10μL to each well, mix well, and place at 37℃ for 5min (if placed at room temperature of 25℃, then 10min). B3.3.2.3 After incubation, pour off the diluted serum, wash 8 times in a row with washing solution, and control dryness. B3.3.2.4 Add the enzyme conjugate diluted according to the working concentration, 0.2mL to each well, and place at 37℃ for 5min. 240
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B3.3.2.5 Pour off the enzyme conjugate, wash 8 times with washing solution, then wash once with distilled water, and drain. B3.3.2.6 Add 3% H,O2 tetramethylbenzidine (TMBS) substrate solution, 0.2mL per well, react for 5-10 minutes and then observe the results.
B3.3.3 Reaction standard:
B3.3.3.1 Visual judgment: judge the results according to the positive control and negative control of each batch. Positive is bright blue, negative is basically colorless. B3.3.3.2 Spectrophotometer colorimetric judgment: do not use HzSO. to stop, then use 590nm wavelength for colorimetric, and judge as positive when P/N≥2.1 (P-patient OD value; N-normal OD value). B4 Latex agglutination test (LA)
B4.1 Dilute the serum sample with physiological saline 1:10. B4.2 Add 1 drop (50μl) of diluted serum and positive and negative control serum to each grid of the reaction plate (black, the same as that used for pregnancy diagnosis latex test).
B4.3 Add 1 drop (50μL) of Schistosoma japonicum latex reagent and shake gently for 10 minutes. Those with clear agglutination are positive, and those without agglutination are negative. B4.4 The positive serum diluted 1:10 can be further diluted in multiples and the above determination is repeated. The highest dilution of serum showing agglutination is the antibody titer.
B5 Monoclonal antibody dot enzyme-linked assay (Dot-ELISA) B5.1 Use a sampling loop to drop the serum to be tested onto the NC fiber membrane (with the hairy surface as the front) and place it at 70℃ for 30 minutes or 37℃ for 2 hours. Place the spotted fiber membrane together with the fiber membranes of the reference positive and reference negative serum in the staining plate. B5.2 Add 200mL of distilled water to reagent No. 1 (PBS) and dissolve it fully to obtain PBS solution. B5.3 Take 150mL of the above PBS solution, add reagent No. 2 (Tween), and mix thoroughly to obtain PBS-Tween solution. Use 20mL of this solution to block and wash the fiber membrane for 30 minutes, and then pour off the solution. B5.4 Take reagent No. 3 (monoclonal antibody enzyme conjugate) and add 10mL of PBS-Tween solution, mix thoroughly, and then add to the staining plate containing the fiber membrane. Place at room temperature for 0.5-1h, shake gently several times in the middle, and pour off the solution. B5.5 Wash the fiber membrane with PBS-Tween solution 3 times, 5 minutes each time, and pour off the solution. B5.6 Add 20ml of PBS solution (without Tween) to reagent No. 4 (substrate), mix well, then add reagent No. 5 (H, O,) and then add to the staining plate, shake gently several times, and pour off the substrate solution after 15 minutes. Wash twice with distilled water to terminate the reaction. The stained fiber membrane is stored in a dark place to dry. B5.7 Carefully observe the color reaction on the fiber membrane and compare it with the spot color reaction presented by the reference serum. The spot color that exceeds the reference negative serum is a positive reaction. According to the intensity of the positive reaction, it can be judged as 10 to 10-10 reactions. Appendix C
(reference)
C1 Acute schistosomiasis
·C1.1 Supportive and symptomatic treatment For severe patients, hydrocortisone or dexamethasone can be added to the rehydration solution and intravenously dripped. After the fever is reduced and the symptoms are improved, pathogen treatment can be started.
C1.2 Pathogenic treatment can be a 6-day therapy with a total dose of 120mg/kg of praziquantel (140mg/kg for children), of which half of the dose is taken on the first and second days, and the remaining half is taken on the third to sixth days. The daily dose is taken in 3 divided doses. C2 Chronic schistosomiasis
Praziquantel 40mg/kg is taken once or twice a day. A total adult dose of 50-60 mg/kg (for children weighing less than 2.11
GB15977-1995
For patients weighing more than 30kg, the total dose is 70mg/kg), 2-day therapy, the daily dose is taken in 2-3 times after meals or between meals, and the dose is still calculated as 60kg for patients weighing more than 60kg. The treatment can be delivered to the door by medical staff, or the medicine can be distributed at designated locations. The elderly, the weak, and those with other diseases should be hospitalized and take medicine at the local schistosomiasis prevention group or health center. During the medication period and within 3-5 days after the end of the medication, they should pay proper attention to rest, reduce physical labor, and avoid high-altitude and water operations.
C3 Late-stage schistosomiasis
C3.1 Pathogen treatment: For most patients with good liver function in the late stage, a total dose of 60mg/kg can be used for 1 or 2 days. For the elderly, the weak, patients with poor liver function or patients with mixed diseases, a total dose of 60mg/kg for 3 days or 90mg/kg for 6 days can be used. The daily dose is divided into 3 doses.
C3.2 Symptomatic treatment
C3.2.1 Splenomegaly: Splenomegaly up to grade I, or splenomegaly up to grade I with obvious hypersplenism, or splenomegaly with esophageal and gastric fundus varices are all indications for spleen surgery. Surgical treatments include simple splenectomy, splenectomy with widened perihilar vascularization, splenectomy with portal azygos vein blood flow occlusion, and splenorenal vein shunt. C3.2.2 Ascites: The treatment principles are the same as those for general cirrhosis ascites. Internal medicine treatment can be combined with traditional Chinese and Western medicine. Western medicine treatment includes supportive therapy, restriction of sodium and water intake, application of nutrition and supportive drugs, and rational use of diuretics. Traditional Chinese medicine uses dialectical treatment. C3.2.3 Upper gastrointestinal bleeding: Internal medicine treatment should be performed first. Including blood volume supplementation, shock correction, rational use of hemostatic agents, intravenous infusion of posterior pituitary hormone and three-lumen balloon packing for hemostasis, and endoscopic injection of sclerosant for hemostasis or gastric lavage with norepinephrine ice water solution can also be considered if necessary. After the bleeding stops and the general condition improves after medical treatment, if there are no contraindications to surgery, elective surgery should be performed in the short term to avoid recurrence of upper gastrointestinal bleeding in the near future. When conservative medical treatment is ineffective, emergency surgery should also be performed decisively and early, such as varicose vein suture, splenectomy and portal azygos vein blood flow blockage or shunt surgery. C3.2.4 Hepatic coma: Comprehensive treatment measures should be adopted, including eliminating the causes of hepatic coma, such as controlling upper gastrointestinal bleeding and clearing intestinal blood accumulation, limiting protein intake, stopping diuretics, correcting water electrolyte and acid-base balance, and taking oral antibiotics to inhibit intestinal bacterial ammonia production. Symptomatic treatment includes reasonable diet and nutrition, the use of drugs to reduce blood ammonia, the use of branched-chain amino acids and levodopa, etc. C3.2.5 Dwarfs: First use praziquantel for pathogen treatment. The dosage can be 70mg/kg for one or two days. The daily dose is divided into three doses. If there is no significant improvement in growth and development about one year after treatment, gonadotropin, thyroxine or gonadotropin can be used for treatment. C4 Indications and contraindications of praziquantel
Praziquantel has a wide range of applications for Japanese schistosomiasis. Acute, chronic and late-stage patients, including schistosomiasis patients with multiple other diseases, generally tolerate well and can complete treatment smoothly. However, clinical experience over the years has shown that cases with severe heart rhythm disorders or uncontrolled heart failure, extremely poor liver compensatory function of late-stage schistosomiasis ascites, and severe renal dysfunction are generally not suitable for treatment. For patients with various types of mental illness and epilepsy, praziquantel treatment should also be extremely cautious, and should generally be carried out in better hospitals, and corresponding measures should be prepared. When patients with cysticercosis are infected with schistosomiasis, praziquantel must be used with caution to treat schistosomiasis. Appendix D
Instructions for the correct use of the standard
(reference)
D1 This standard applies to patients found in schistosomiasis endemic areas through surveys [only the COPT ring sedimentation rate can be relaxed to 1%) and to patients diagnosed with schistosomiasis in hospitals and clinics, and to staging and etiological treatment of patients. D2 The serological diagnostic method used to diagnose patients must use relatively standardized antigens or test kits. D3 Drug treatment: Use praziquantel tablets. 242
Additional instructions:
GB15977-1995
This standard was proposed by the Ministry of Health of the People's Republic of China. This standard was drafted by the Institute of Parasitic Diseases, Chinese Academy of Preventive Medicine. The main drafters of this standard are Qiu Lishu and Chen Minggang. This standard is entrusted by the Ministry of Health to the Office of Infectious Disease Supervision and Management of the Ministry of Health for interpretation. 2.13
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