GB 15985-1995 Diagnostic criteria and treatment principles for filariasis
Some standard content:
National Standard of the People's Republic of China
Diagnostic criteria and principles of management of filariasis
GB15985-1995
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. 1 Subject matter and scope of application
This standard specifies the diagnostic criteria, treatment methods and prevention and control principles for each stage of filariasis (Bancroftian filariasis and Malayan filariasis), namely microfilariasis, acute filariasis and chronic filariasis. This standard is applicable to the prevention and control of filariasis by professional institutions in epidemic areas and the diagnosis and treatment of filariasis patients by health and epidemic prevention and medical care institutions at all levels.
2 Diagnostic principles
Diagnosis is made based on the residence history, clinical manifestations, etiological examination, serum immunological examination, etc. in the epidemic season in the filariasis endemic area. 3 Diagnostic criteria
3.1 Microschisis
3.1.1 History of residence in endemic areas during the epidemic season.
3.1.2 Microschisis positive in nighttime blood draw. Confirmed cases: meet 3.1.2.
3.2 Acute filariasis
3.2.1 History of residence in endemic areas during the epidemic season.
3.2.2 Recurrent non-bacterial infectious limb (or scrotal, female breast) lymphadenitis/lymphangitis (or spermatic cord inflammation, orchitis, epididymitis), peripheral pain, tenderness, swelling, warmth, or erysipelas-like dermatitis, symptoms lasting more than 3 days, accompanied by systemic symptoms such as fever, headache, and malaise.
Note: Acute inflammation of Malayan filariasis is limited to the limbs. 3.2.3 Microschisis positive in nighttime blood draw. 3.2.4 Antibody positive in indirect fluorescent antibody test or enzyme-linked immunosorbent assay. Suspected cases: meet 3.2.1 and 3.2.2. Confirmed cases: Suspected cases plus 3.2.3 or suspected cases plus 3.2.4. 3.3 Chronic filariasis
3.3.1 Long-term history of living in endemic areas.
3.3.2 Asymmetric limb lymphedema, elephantiasis, hydrocele, chyluria, and scrotal or female breast enlargement (chronic signs of Malayan filariasis are limited to limb lymphedema and elephantiasis, and the swelling is limited to the distal end of the knee and elbow joints). Or have the manifestations of 3.2.2. 3.3.3 Positive microfilaments in night blood test. Approved by the State Administration of Technical Supervision on December 15, 1995, and implemented on July 1, 1996
GB15985-1995
3.3.4 Positive antibodies detected by indirect fluorescent antibody test or enzyme-linked immunosorbent assay. 3.3.5 Microfilaments are found in urine, lymphatic fluid, hydrocele (or other aspirated fluids), adult worms are found in lymphatic vessels and lymph nodes, or filarial sections are found in pathological tissue sections.
Suspected case: meets 3.3.1 and 3.3.2. Confirmed cases: Suspected cases should add 3.3.5 or 3.3.3 or 3.3.4. 4 Treatment principles
4.1 Pathogen treatment
4.1.1 Individual treatment
4.1.1.1 Bancroftian filariasis
Generally, a total dose of 4.2g of diethylcarbamazine is used for 7 days (total amount 70-84mg/kg), that is, 0.2g each time, 3 times a day, for 7 consecutive days (adult dosage, children's dosage should be reduced, pregnant women, lactating women and patients with serious diseases should delay treatment or exempt from treatment, the same below) as one course of treatment, and retreatment is required for 2-3 courses, with an interval of more than half a month. 4.1.1.2 Malayan filariasis
Generally, a 4-day or 2-day treatment with a total amount of 2g of Haiqunsheng (total amount 33~40mg/kg) is used, that is, 0.5g taken at a time for 4 consecutive days or 0.5g twice a day for 2 consecutive days, and retreatment for 2~3 courses of treatment, with an interval of more than half a month. For patients with a high density of microfilaments, the first treatment should be a 10-day micro-incremental therapy of Haiqunsheng 1g (that is, on the 14th day, 12.5mg per day, on the 5th to 7th day, 25mg per day, on the 8th to 9th day, 50mg per day, and 1.0g taken at a time on the 10th day).
4.1.2 Group treatment
4.1.2.1 Bancroftian filariasis
4.1.2.1.1 In areas where the prevalence of microfilaments in the population is 10% to 20%, after a blood test survey, the whole population shall be given 0.3% sea quinoline salt for 6 months (the total amount of sea quinoline is about 9g), and those with microfilaments shall be given 3g of sea quinoline for 5 days, that is, 0.3g each time, twice a day, 5 days as a course of treatment, 2 to 3 courses of treatment, with an interval of more than half a month. 4.1.2.1.2 In areas where the prevalence of microfilaments in the population is 5% to 10%, after a blood test group sampling survey, the whole population shall be given 0.3% sea quinoline salt for 6 months.
4.1.2.1.3 In areas where the prevalence of microfilaments in the population is below 5%, a general survey shall be conducted. For patients with microfilaments, 3g of Haiqunsheng shall be used for 5 days for 2 to 3 courses of treatment, with an interval of more than half a month.
4.1.2.2 Filariasis in chickens
4.1.2.2.1 In areas where the prevalence of microfilaments in the population is between 5% and 10%, a general survey shall be conducted. For patients with microfilaments, 2g of Haiqunsheng shall be used for 4 or 2 days for 3 to 4 courses of treatment, with an interval of more than half a month. It is also possible to adopt a universal administration of 0.3% Haiqunsheng medicinal salt for 4 to 6 months. 4.1.2.2.2 In areas where the prevalence of microfilaments in the population is below 5%, a general survey shall be conducted. For patients with microfilaments, 2g of Haiqunsheng shall be used for 4 or 2 days for 2 to 3 courses of treatment, with an interval of more than half a month. 4.1.2.2.3 The best time for group treatment is in winter and spring. 4.2 Symptomatic treatment (see Appendix C for details)
4.3 Vector control
4.3.1 In conjunction with environmental improvement and new rural construction, eliminate mosquito breeding grounds. 4.3.2 Promote the use of mosquito nets, screen windows, screen doors and other mosquito-proof equipment and the use of mosquito repellents. 4.3.3 In areas where Malayan filariasis is prevalent where Anopheles anthropophagus is present, insecticides can be used to kill mosquitoes indoors in conjunction with filariasis control. 294
A1 Blood test
A1.1 Blood film method
GB15985—1995
Appendix A
Pathological examination
(Supplement)
A1.1.1 Preparation of blood film: From 9 pm to 2 am the next morning, disinfect the earlobe (or fingertip) with an alcohol cotton ball and wait for it to dry. Use a three-edged needle to quickly and deeply puncture and gently squeeze out the blood. Take 3 large drops of blood, about 60uL, and place them on a numbered clean slide. Smear into an oval or rectangular thick blood film (about 3cm long and 1.5cm wide) with neat edges and uniform thickness, and place it flat in a covered slide box to prevent dust or insects from eating. A1.1.2 Blood film staining: The next day, put the naturally dried blood film into clean water for hemolysis for 5~10 minutes until the blood film becomes milky white, take it out to dry, fix it with methanol, and stain it. Borax-methylene blue staining can be used for large-scale surveys. Giemsa solution or hematoxylin staining is suitable for identification of insect species and preservation. A1.1.2.1 Borax-methylene blue staining method: Take 2g of methylene blue and 3g of borax, put them in a mortar, add water while grinding, rinse into a bottle after dissolving, add 100mL of distilled water to make the stock solution, filter and set aside. When staining, take 5mL of the stock solution and add clean water to make a 5% dilution, stain for 3 to 5 minutes, so that the blood film is sky blue, and then rinse gently with clean water. This method does not require hemolysis and fixation before staining. A1.1.2.2 Giemsa staining method: The dye solution is made of 0.5g Giemsa powder, 25mL of neutral glycerol and 25mL of methanol. First, place the dye powder in a mortar, add a small amount of glycerol to fully grind, add while grinding, until the glycerol is added, then pour it into a 100mL glass stopper bottle, wash it with a small amount of methanol several times, pour the glycerol dye solution into the bottle, plug the bottle tightly, shake it thoroughly, and place it in a 55-60℃ incubator for 24 hours or at room temperature for 3-5 days. This is the stock solution. The longer it is placed, the better the dyeing effect. Dilute it before use. Fix the blood smear that has dried after hemolysis with methanol for about 1 minute, then add 10% dye solution (the stock solution is diluted with clean water) and dye for 45 minutes, and rinse gently with distilled water. A1.1.2.3 Debrie's hematoxylin staining method, the dye solution is made up of A solution, i.e. saturated ammonium alum solution (20g ammonium alum, 100mL distilled water); B solution (1g hematoxylin crystals plus 10mL pure alcohol), and C solution (25mL glycerol plus 25mL methanol). Add liquid B dropwise into liquid A, pour into a container without a tight lid, place in an airy place, allow to fully oxidize for 2 weeks to 1 month, filter and add liquid C, seal and set aside. The steps for hemolysis and fixation of blood smears are the same as before. Dye with the above dye solution for 10~20 minutes, decolorize with 0.05%~0.1% dilute hydrochloric acid for a while, and rinse with running water until the blood film turns blue. A1.1.3 Microscopic examination of blood smears: Check and count the microfilaments in the stained blood smears in sequence under a low-power microscope, and identify them based on the size, body shape, refractive index, presence or absence of sheath, whether the epidermis is smooth, and internal structure of the microfilaments. Observe the body core and head end space, nerve ring, excretory cells, excretory pores, anal pores, tail nucleus and other structures of the microfilaments under a high-power microscope. If necessary, use an oil immersion lens for further identification. A1.1.4 The key points for distinguishing the morphology of Benedict's and Malayan microfilaments are shown in Table A1. Table A1
Length, um
Width·μm
Gap at the head end
(length-width ratio)
A1.2 Microfilament concentration method
Benedict's microfilament
244～296 (average 260)
Soft, natural bend, no small bend
(1 11)
shaped, neatly arranged, each nucleus is separated, clearly countable, without
malayan microfilaments
177~230 (average 220)
rigid, with small bends on the large bend
oval, different sizes, closely arranged, often overlapped, difficult to distinguish two
A1.2.1 Improved distilled water method: Take 1mL of venous blood (the time of blood collection is the same as A1.1), place it in a centrifuge containing 0.4mL 3.8% sodium succinate In the heart tube, add 8-10mL of distilled water after mixing, shake repeatedly, centrifuge at 3000 rpm for 3-5min after the red blood cells are dissolved, pour off the upper liquid, add 8-10mL of 0.05mol/L sodium hydroxide, press the tube mouth, shake vigorously several times, and leave for 5-10min to dissolve the fibrin clot quickly, centrifuge again, remove the supernatant, smear the sediment, wait to dry, stain, and examine under a microscope. A1.2.2 Microporous membrane filtration method: rinse the numbered microporous membrane with a diameter of 25mm and a pore size of 5um with distilled water and put it into the filter, and place a filter paper of the same size moistened with physiological saline under the filter membrane. Use a syringe to take 1mL of venous blood (the time of blood collection is the same as A1.1), add 0.1mL of 5% sodium citrate for anticoagulation, inhale 9mL of 10% sodium sulfate of polyoxyethylene fatty alcohol, or 2% Tween-80, 0.1% sodium bicarbonate or 10% sodium sulfate of polyoxyethylene fatty alcohol (ES) solution, mix well, remove the needle, insert it directly into the filter, slowly push the syringe to make the hemolyzed blood pass through the filter membrane, and wash the filter membrane 3 times with 10mL of normal saline. Open the filter, take out the film, dry it naturally, stain it in hot hematoxylin for 5min, wash it with water, wait to dry, and examine it under a microscope.
A2 Examination of microfilaments in lymph fluid, hydrocele, and chylous urine A2.1 Lymph fluid, hydrocele (or other extracted fluid): directly smear or dilute it 10 times with normal saline and centrifuge it to check the sediment. If the liquid protein content is high and it is gelatinous and easy to coagulate, add anticoagulant and check it. A2.2 Chylous urine (or chylous effusion): Take 4 mL of chylous urine in a test tube, add 2 mL of ether, mix and shake to fully dissolve the fat in the chyle, remove the upper fat, dilute 10 times with water, and centrifuge for examination. A3 Biopsy
A3.1 Check for adult worms in lymphatic vessels and lymph nodes: Fix the nodules removed by surgery on a wooden board or cork board with a pin, separate the tissue around the nodules, carefully cut the diseased lymphatic wall, separate the contents, and take out the caseous pus for examination. If the nodules are removed and dissected 2 weeks after the appearance of the nodules, the pus in the tube may have fibrosis. The pus contains a large number of macrophages, monocytes, eosinophils and Xia Lei crystals. Move the caseous substance or fibrous tissue to a glass slide, add a few drops of physiological saline, and then separate the outer tissue with a dissecting needle to see the surrounded adult worms. If the time is too long, adhesion will occur and the worm body will be broken into pieces, which makes it difficult to separate the worm body from the tissue. In this case, one needle can be used to fix the surrounding tissue, and another needle can be used to gently move the worm body along its long axis to one end, or the tissue can be moved to culture blood containing physiological saline to make the worm body half-floating and easier to separate. However, if the nodule is formed for a long time, it is difficult to obtain a complete adult worm. The adult worm or worm segment can be stored in glycerol alcohol (70% alcohol 95mL, glycerol 5mL) for identification.
A3.2 Histopathological examination: Cut off the suspicious lymph nodes, lymphatic nodules or other tissues, fix them with 10% neutral formaldehyde for 1 to 2 days, and move them to 70% alcohol for pathological sections. In addition to adult filarial worms, pathological changes such as eosinophils, reticuloendothelial cells, lymphocytes, plasma cells, fibroblasts, foreign body giant cells, granulomas, pseudotuberculosis, fibrous tissue hyperplasia, lymphangiitis, and intraluminal granulation tissue hyperplasia forming emboli or polyp-like obstruction of the lumen can be seen in the sections. In the sections, if the microfilament nuclei in the female worm with clear structure are clear, there are a small amount of plasma cells, lymphocytes and eosinophils infiltrating the lymphatic vessel wall, and there is no cellulose deposition or granuloma formation, then the adult worm may be alive. If there are more eosinophils and scattered tissues around the worm body, the worm body is surrounded by inflammatory cells to form granulomas, fibrotic nodules are formed after fibrosis, or the worm body in the nodule is calcified, then they are all dead worms. If there is neutrophil infiltration and exudation, and the intimal endothelial cells are swollen and hyperplastic, it is a secondary bacterial infection.
Appendix B
Serum immunological examination
(Supplement)
B1 Indirect fluorescent antibody test (IFAT)
B1.1 Antigen slice
B1.1.1 Frozen section antigen of Brugia malayi adults: Infect long-clawed gerbils with larvae of the infective stage of Brugia malayi. Six months later, collect female adults from the abdominal cavity of infected mice. Wash twice with saline, rinse once with distilled water, embed in 3% to 5% methylcellulose, and cut into 45μm thick sections (each section contains 4 to 6 worm cross sections) with a freezing section machine, attach to a clean slide, fix with acetone, dry and store at -20℃ for later use.
GB15985--1995
B1.1.2 Malayan microfilament fragment antigen: Collect Malayan microfilaments from the peritoneal fluid of long-clawed gerbils infected with Malayan filariasis and wash with PBS (or saline). The suspension is placed in an ice cube overnight, concentrated by centrifugation the next day, fixed with methanol, and then repeatedly ultrasonically broken. The output power of the ultrasonic crusher is not less than 25W (30s each time, 30-60s interval, 3-5min in total), until the microfilament fragments are 20-30μm long. Drop 1 drop of microfilament fragment suspension (≥100 fragments/drop) on a clean glass slide, heat-dry at about 50℃ for 3min before use. B1.2 Blood sample: serum or filter paper blood.
B1.2.1 Serum: Collect venous blood or peripheral blood from the subject, collect serum after centrifugation. Transport at low temperature, store at 4℃, and store at -20℃ for about 2 years. Do not freeze and thaw repeatedly during this period to avoid affecting the antibody titer. B1.2.2 Filter paper dried blood drop: Use quantitative Xinhua filter paper to collect blood from the earlobe or fingertip of the subject, so that the blood drop diameter is 1.2cm and the blood volume is about 20μL (equivalent to 10μL of serum). After numbering and drying, put it in a plastic bag with desiccant and store it at low temperature. It can be stored for half a year at 4℃ and for 2 years at -20℃.
B1.3 Operation method: Dilute the test serum with pH8.0, 0.01mol/LPBS to 1:10 or 1:20 (each filter paper dried blood drop is soaked in 0.1 or 0.2ml PBS solution for 30min). Take out the antigen slice from the refrigerator, and after rewarming, add 1 drop of the diluted clear to be tested on each slice or drop of antigen, and incubate at 37℃ for 30min. Wash with PBS 3 times (5 minutes each time), blow dry with cold air, add goat anti-human IgG fluorescent antibody diluted with PBS, incubate as above, wash, use 0.01% Evans blue as background stain for 2 minutes, wash as above, blow dry. Add buffered glycerol (pH8.0) and examine with coverslip under fluorescence microscope. Blood samples with positive reaction need to be continuously diluted and tested until negative reaction. The dilution of the last positive reaction is the positive endpoint titer. B1.4 Reaction standard
B1.4.1 The section antigen of Brugia malayi adult worm is divided into 4 levels according to the fluorescence intensity and fluorescence coloring range of the worm body section: "two" worm body section star orange red, no obvious yellow-green fluorescence; "ten" yellow-green fluorescence of average brightness, with a limited color range; "ten-ten" yellow-green fluorescence of medium brightness, with a wide range of color development; "ten-ten+" bright yellow-green fluorescence, with a wide range of color development. Generally, a positive result is determined when the fluorescence reaction intensity is "10" or above at a blood sample titer of 1:20. B1.4.2 Microfilament fragment antigens are divided into four levels according to the fluorescence intensity and coloring range of microfilament fragments: "--" The microfilament fragments are fuzzy light yellow-green or red; "10" The fragments are yellow-green fluorescent, shaped like dumbbells, and the red color is obvious; "10+10" The fragments are significantly yellow-green fluorescent, and the membrane also has yellow-green fluorescence; "+10" The fragments and membranes are bright yellow-green fluorescent. Generally, a positive result is determined when the fluorescence reaction is "10" or above at a blood sample titer of 1:10. B2 Enzyme-linked immunosorbent assay (ELISA)
B2.1 Antigen: Soluble antigen of adult Brugia malayi. Adult Brugia malayi are collected from the abdominal cavity of long-clawed gerbils infected with Brugia malayi. After the worm bodies are washed with physiological saline and rinsed with distilled water, they are defatted and dried with acetone, and then 0.0.1% thimerosal physiological saline was ground into a homogenate, frozen and thawed for 3 days (twice a day), and then cold soaked at 4°C for 3-4 days. After ultrasonic crushing, centrifuged at low temperature and high speed (7000r/min) for 30 minutes. The supernatant is the soluble antigen. The optical density values at 235 and 280nm wavelengths were measured by spectrophotometer ()D, and the amount of antigen protein was calculated according to formula (B1): X (mg/mL) (OD235-OD280) X0.4.... B2.2 Operation method
· (B1)
B2.2.1 Conventional indirect ELISA: Dilute the antigen to an appropriate working concentration (2-5μg/well) with pH9.6, 0.05mol/1 carbonate buffer, coat the polystyrene reaction plate, add 0.3mL to each well, and incubate at 4°C overnight. The next day, the antigen solution was poured off, washed 3 times with pH7.2PBS/Tween-20 (PBS/T), 5 minutes each time, and dried. The serum to be tested (or blood droplets dried on filter paper) was diluted to 1:200 with PBS/T, 0.3 mL was added to each well, 3 wells were added to each sample, and the slide box was incubated at 37C for 2 hours, washed as above, and dried. 0.3 mL of horseradish peroxidase-labeled goat anti-human IgG in PBS/T was added to each well, incubated as above, washed, and dried. Then, the substrate solution [40 mg of o-phenylenediamine, 50 mL of citric acid-phosphate buffer, 50 ml of distilled water, and 40 μL of 30% hydrogen peroxide (H2O2) and 0.3 mL were added, and incubated as above. Finally, 75 μL of 2 mol/L sulfuric acid (HSO2) was added to each well to terminate the reaction. The liquid matrix of 3 wells of each sample was aspirated into a colorimetric cup (cup diameter 1 cm), and the optical density (OD) value at a wavelength of 492 nm was measured on a spectrophotometer. Each plate was set with positive reference serum, negative reference serum, and blank control wells. The measured sample OD value was corrected. Sample OD value minus control well OD value
calibrated sample OD value = positive reference serum OD value minus control well OD value (B2)
GB15985-1995
B2.2.2 Improved indirect ELISA: dilute the antigen with 0.05 mol/L carbonate buffer at pH 9.6 at an appropriate working concentration, coat a 40-well polystyrene reaction plate, add 0.1 mL to each well, and incubate at 4°C overnight. Pour off the antigen solution the next day, wash 3 times with pH 7.2 PBS/T, 5 min each time, and spin dry. Dilute the blood sample to be tested with PBS/T to 1:200, add 0.1 mL to each well, add 2 wells to each sample, place in a humidified box, incubate at 37°C for 30 min, wash as above, and spin dry. Add 0.1 mL of horseradish peroxidase-labeled goat anti-human IgG diluted with PBS/T to each well, incubate as above, wash, and spin dry. Then add 0.1mL of substrate solution, incubate as above, and add 25μL of 2mol/L sulfuric acid (H2SO4) to terminate the reaction. Use a special enzyme label detector (after correcting the zero point with the PBS control well) to measure the OD value of each well at a wavelength of 492 (or 490)nm. Each plate has a positive reference serum, a negative reference serum and a PBS control. The OD value of each sample is calculated as the average OD value of the two wells, and then corrected with the OD value of the positive reference serum on the same plate. Sample OD value
Correction sample OD value one hundred
Positive reference serum OD value
B2.3 Reaction standard
（B3）
Measure more than 100 healthy human sera, calculate their OD values and standard deviations (SD), and use the OD mean plus 2SD as the positive reaction threshold (usually 0.4). When the sample OD value is equal to or greater than this value, it is judged as positive. Or the ratio of the sample OD value (P) to the negative reference serum OD value (N) P/N≥2 is positive.
Preparation of B3 solution
B3. 1 pH8. 0, 0. 01mol/L PBS solution stock solution 0.2mol/L phosphate buffer solution
a. NaH2PO4 · 2H,O
distilled water added to
b. Na2HPO,·12H,O
distilled water added to
Take 5.3mL of solution a, 94.7mL of solution b, 17g of NaCl and add water to 2000mL. B3.2 pH8.0 buffered glycerol
5 parts of glycerol (first grade) and 1 part of 0.01mol/LPBS (pH8.0) are mixed and stored in the refrigerator (no bubbles). B3.3pH9.6, 0.05mol/L carbonate buffer Na2CO3
NaHCO3
Add distilled water to
B3.4pH7.2PBS/Tween-20
Na2HPO, : 12H20
Tween-20
Add distilled water to
1000mL
1 000 mL
B3.5pH5.0 phosphate-citrate buffer substrate solution 0.1mol/L citric acid solution (21.041g/L) 0. 2mol/L Na2HPO, (28. 4g/L)
Distilled water
o-phenylenediamine
Add 30% hydrogen peroxide (H2O2) before use
40 μL
C1 Acute filariasis (limbs, scrotum, female breast) GB15985—1995
Symptomatic treatment
(reference)
Use anti-inflammatory analgesics, and stop taking them after acute symptoms are relieved. For patients with acute lymphadenitis and lymphangitis (flushing) of the lower limbs, if anti-inflammatory analgesics can be taken during the premonitory period of the attack, the acute symptoms can be significantly alleviated or the attack can be stopped. Patients with bacterial infection need to be given antibacterial treatment. C2 Chronic filariasis
C2.1 Limb lymphedema and elephantiasis
Baking and bandaging therapy: Use radiant heat or microwave diathermy to bake the affected limb and then bandage it with an elastic bandage. Once a day, the former is 1 hour each time, 20 times as a course of treatment, rest for half a month, and then proceed to the next course of treatment; the latter is 30 minutes each time, 15 times as a course of treatment, rest for 2 months, and then proceed to the next course of treatment. During the baking treatment and rest period, the affected limb should be continuously bandaged with an elastic bandage during the day for 2 to 3 courses of treatment. For patients with tinea pedis, antifungal treatment is used to control fungal infection. C2.2 Chylous urine
-General treatment: Pay attention to rest during the attack, and avoid eating oily foods, meat and eggs. For those with chylous clots, dysuria and urinary retention, the amount of water consumed should be reduced, and the lower abdomen should be massaged with hands; or medium-chain triglycerides (MCT) can be used instead of ordinary edible oils to quickly relieve the patient's pain. The dosage of MCT is 4 to 5g per serving for adults, 3 times a day, and the course of treatment is 1 month. It can be taken at intervals of 2 to 3 courses. Frequent attacks and severe cases can be treated with silver nitrate infusion or surgery. C2.3 Hydrocele
Surgical treatment: Hydrocele with a large amount of hydrocele is treated with hydrocele flipping. Additional notes:
This standard is proposed by the Ministry of Health of the People's Republic of China. This standard is drafted by the Institute of Parasitic Diseases, Chinese Academy of Preventive Medicine. The main drafters of this standard are Sun Dejian, Shi Zongjun and Deng Shanshan. The Ministry of Health has entrusted the technical coordination unit, the Office of Supervision and Administration of Communicable Disease Prevention and Control of the Ministry of Health, to interpret this standard.1mL, 4℃ overnight. The next day, the antigen solution was removed, and the plate was washed with pH7.2PBS/T for 3 times, 5min each time, and then dried. The blood sample was diluted with PBS/T to 1:200, 0.1mL was added to each well, and 2 wells were added to each sample. The plate was placed in a humidified box, incubated at 37℃ for 30min, washed as above, and dried. 0.1mL of horseradish peroxidase-labeled goat anti-human IgG diluted with PBS/T was added to each well, incubated as above, washed, and dried. Then 0.1mL of substrate solution was added, and after incubation as above, 25μL of 2mol/L sulfuric acid (H,SO,) was added to terminate the reaction. The OD value of each well at a wavelength of 492 (or 490)nm was measured using a dedicated enzyme label detector (after calibrating the zero point with the PBS control well). Each plate was set up with a positive reference serum, a negative reference serum, and a PBS control. The OD value of each sample was calculated as the average OD value of the 2 wells, and then corrected with the OD value of the positive reference serum on the same plate. Sample OD value
Correction sample OD value one hundred
Positive reference serum OD value
B2.3 Reaction standard
（B3）
Measure more than 100 healthy people's sera, calculate their OD value and standard deviation (SD), and use the OD mean plus 2SD as the positive reaction threshold (usually 0.4). When the sample OD value is equal to or greater than this value, it is judged as positive. Or when the ratio of the sample OD value (P) to the negative reference serum OD value (N) P/N ≥ 2 is positive.
Preparation of B3 solution
B3. 1 pH8. 0, 0. 01mol/L PBS solution stock solution 0.2mol/L phosphate buffer solution
a. NaH2PO4 · 2H,O
distilled water added to
b. Na2HPO,·12H,O
distilled water added to
Take 5.3mL of solution a, 94.7mL of solution b, 17g of NaCl and add water to 2000mL. B3.2 pH8.0 buffered glycerol
5 parts of glycerol (first grade) and 1 part of 0.01mol/LPBS (pH8.0) are mixed and stored in the refrigerator (no bubbles). B3.3pH9.6, 0.05mol/L carbonate buffer Na2CO3
NaHCO3
Add distilled water to
B3.4pH7.2PBS/Tween-20
Na2HPO, : 12H20
Tween-20
Add distilled water to
1000mL
1 000 mL
B3.5pH5.0 phosphate-citrate buffer substrate solution 0.1mol/L citric acid solution (21.041g/L) 0. 2mol/L Na2HPO, (28. 4g/L)
Distilled water
o-phenylenediamine
Add 30% hydrogen peroxide (H2O2) before use
40 μL
C1 Acute filariasis (limbs, scrotum, female breast) GB15985—1995
Symptomatic treatment
(reference)
Use anti-inflammatory analgesics, and stop taking them after acute symptoms are relieved. For patients with acute lymphadenitis and lymphangitis (flushing) of the lower limbs, if anti-inflammatory analgesics can be taken during the premonitory period of the attack, the acute symptoms can be significantly alleviated or the attack can be stopped. Patients with bacterial infection need to be given antibacterial treatment. C2 Chronic filariasis
C2.1 Limb lymphedema and elephantiasis
Baking and bandaging therapy: Use radiant heat or microwave diathermy to bake the affected limb and then bandage it with an elastic bandage. Once a day, the former is 1 hour each time, 20 times as a course of treatment, rest for half a month, and then proceed to the next course of treatment; the latter is 30 minutes each time, 15 times as a course of treatment, rest for 2 months, and then proceed to the next course of treatment. During the baking treatment and rest period, the affected limb should be continuously bandaged with an elastic bandage during the day for 2 to 3 courses of treatment. For patients with tinea pedis, antifungal treatment is used to control fungal infection. C2.2 Chylous urine
-General treatment: Pay attention to rest during the attack, and avoid eating oily foods, meat and eggs. For those with chylous clots, dysuria and urinary retention, the amount of water consumed should be reduced, and the lower abdomen should be massaged with hands; or medium-chain triglycerides (MCT) can be used instead of ordinary edible oils to quickly relieve the patient's pain. The dosage of MCT is 4 to 5g per serving for adults, 3 times a day, and the course of treatment is 1 month. It can be taken at intervals of 2 to 3 courses. Frequent attacks and severe cases can be treated with silver nitrate infusion or surgery. C2.3 Hydrocele
Surgical treatment: Hydrocele with a large amount of hydrocele is treated with hydrocele flipping. Additional notes:
This standard is proposed by the Ministry of Health of the People's Republic of China. This standard is drafted by the Institute of Parasitic Diseases, Chinese Academy of Preventive Medicine. The main drafters of this standard are Sun Dejian, Shi Zongjun and Deng Shanshan. The Ministry of Health has entrusted the technical coordination unit, the Office of Supervision and Administration of Communicable Disease Prevention and Control of the Ministry of Health, to interpret this standard.1mL, 4℃ overnight. The next day, the antigen solution was removed, and the plate was washed with pH7.2PBS/T for 3 times, 5min each time, and then dried. The blood sample was diluted with PBS/T to 1:200, 0.1mL was added to each well, and 2 wells were added to each sample. The plate was placed in a humidified box, incubated at 37℃ for 30min, washed as above, and dried. 0.1mL of horseradish peroxidase-labeled goat anti-human IgG diluted with PBS/T was added to each well, incubated as above, washed, and dried. Then 0.1mL of substrate solution was added, and after incubation as above, 25μL of 2mol/L sulfuric acid (H,SO,) was added to terminate the reaction. The OD value of each well at a wavelength of 492 (or 490)nm was measured using a dedicated enzyme label detector (after calibrating the zero point with the PBS control well). Each plate was set up with a positive reference serum, a negative reference serum, and a PBS control. The OD value of each sample was calculated as the average OD value of the 2 wells, and then corrected with the OD value of the positive reference serum on the same plate. Sample OD value
Correction sample OD value one hundred
Positive reference serum OD value
B2.3 Reaction standard
（B3）bZxz.net
Measure more than 100 healthy people's sera, calculate their OD value and standard deviation (SD), and use the OD mean plus 2SD as the positive reaction threshold (usually 0.4). When the sample OD value is equal to or greater than this value, it is judged as positive. Or when the ratio of the sample OD value (P) to the negative reference serum OD value (N) P/N ≥ 2 is positive.
Preparation of B3 solution
B3. 1 pH8. 0, 0. 01mol/L PBS solution stock solution 0.2mol/L phosphate buffer solution
a. NaH2PO4 · 2H,O
distilled water added to
b. Na2HPO,·12H,O
distilled water added to
Take 5.3mL of solution a, 94.7mL of solution b, 17g of NaCl and add water to 2000mL. B3.2 pH8.0 buffered glycerol
5 parts of glycerol (first grade) and 1 part of 0.01mol/LPBS (pH8.0) are mixed and stored in the refrigerator (no bubbles). B3.3pH9.6, 0.05mol/L carbonate buffer Na2CO3
NaHCO3
Add distilled water to
B3.4pH7.2PBS/Tween-20
Na2HPO, : 12H20
Tween-20
Add distilled water to
1000mL
1 000 mL
B3.5pH5.0 phosphate-citrate buffer substrate solution 0.1mol/L citric acid solution (21.041g/L) 0. 2mol/L Na2HPO, (28. 4g/L)
Distilled water
o-phenylenediamine
Add 30% hydrogen peroxide (H2O2) before use
40 μL
C1 Acute filariasis (limbs, scrotum, female breast) GB15985—1995
Symptomatic treatment
(reference)
Use anti-inflammatory analgesics, and stop taking them after acute symptoms are relieved. For patients with acute lymphadenitis and lymphangitis (flushing) of the lower limbs, if anti-inflammatory analgesics can be taken during the premonitory period of the attack, the acute symptoms can be significantly alleviated or the attack can be stopped. Patients with bacterial infection need to be given antibacterial treatment. C2 Chronic filariasis
C2.1 Limb lymphedema and elephantiasis
Baking and bandaging therapy: Use radiant heat or microwave diathermy to bake the affected limb and then bandage it with an elastic bandage. Once a day, the former is 1 hour each time, 20 times as a course of treatment, rest for half a month, and then proceed to the next course of treatment; the latter is 30 minutes each time, 15 times as a course of treatment, rest for 2 months, and then proceed to the next course of treatment. During the baking treatment and rest period, the affected limb should be continuously bandaged with an elastic bandage during the day for 2 to 3 courses of treatment. For patients with tinea pedis, antifungal treatment is used to control fungal infection. C2.2 Chylous urine
-General treatment: Pay attention to rest during the attack, and avoid eating oily foods, meat and eggs. For those with chylous clots, dysuria and urinary retention, the amount of water consumed should be reduced, and the lower abdomen should be massaged with hands; or medium-chain triglycerides (MCT) can be used instead of ordinary edible oils to quickly relieve the patient's pain. The dosage of MCT is 4 to 5g per serving for adults, 3 times a day, and the course of treatment is 1 month. It can be taken at intervals of 2 to 3 courses. Frequent attacks and severe cases can be treated with silver nitrate infusion or surgery. C2.3 Hydrocele
Surgical treatment: Hydrocele with a large amount of hydrocele is treated with hydrocele flipping. Additional notes:
This standard is proposed by the Ministry of Health of the People's Republic of China. This standard is drafted by the Institute of Parasitic Diseases, Chinese Academy of Preventive Medicine. The main drafters of this standard are Sun Dejian, Shi Zongjun and Deng Shanshan. The Ministry of Health has entrusted the technical coordination unit, the Office of Supervision and Administration of Communicable Disease Prevention and Control of the Ministry of Health, to interpret this standard.2 Chylous urine
-General treatment: During the attack, pay attention to rest, avoid eating oily foods, meat and eggs. Patients with chylous clots, dysuria and urinary retention should reduce water intake and massage the lower abdomen with hands; or use medium-chain triglycerides (MCT) instead of ordinary edible oils to quickly relieve the patient's pain. The dosage of MCT for adults is 4-5g each time, 3 times a day, and the course of treatment is 1 month. It can be taken at intervals for 2-3 courses. Frequent attacks and severe cases can be treated with silver nitrate infusion or surgery. C2.3 Hydrocele
Surgical treatment: Hydrocele is used for large amounts of hydrocele. Additional notes:
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 Sun Dejian, Shi Zongjun and Deng Shanshan. This standard is interpreted by the Ministry of Health's Office of Supervision and Administration of Communicable Disease Prevention and Control, the technical unit entrusted by the Ministry of Health. 2992 Chylous urine
-General treatment: During the attack, pay attention to rest, avoid eating oily foods, meat and eggs. Patients with chylous clots, dysuria and urinary retention should reduce water intake and massage the lower abdomen with hands; or use medium-chain triglycerides (MCT) instead of ordinary edible oils to quickly relieve the patient's pain. The dosage of MCT for adults is 4-5g each time, 3 times a day, and the course of treatment is 1 month. It can be taken at intervals for 2-3 courses. Frequent attacks and severe cases can be treated with silver nitrate infusion or surgery. C2.3 Hydrocele
Surgical treatment: Hydrocele is used for large amounts of hydrocele. Additional notes:
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 Sun Dejian, Shi Zongjun and Deng Shanshan. This standard is interpreted by the Ministry of Health's Office of Supervision and Administration of Communicable Disease Prevention and Control, the technical unit entrusted by the Ministry of Health. 299
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