This standard specifies the diagnosis and treatment principles of cystic echinococcosis and alveolar echinococcosis. It is applicable to the diagnosis and treatment of both types of echinococcosis by medical and health institutions at all levels and of all types, and for endemic disease prevention and control institutions. GB 17013-1997 Diagnostic Criteria and Treatment Principles for Echinococcosis GB17013-1997 Standard Download Decompression Password: www.bzxz.net

【GB170131997】
Diagnostic criteria and treatment principles for echinococcosis
Echinococcosis, also known as echinococcosis, is a disease caused by the larvae of Echinococcus parasitizing the human body. There are two types in my country, namely cystic echinococcosis caused by Echinococcus granulosus and alveolar echinococcosis caused by Echinococcus multilocularis. There are local cystic echinococcosis patients in 22 provinces (municipalities) and autonomous regions in my country. Cases of alveolar echinococcosis have been reported in Ningxia, Xinjiang, Gansu, Sichuan, Qinghai, Tibet and Heilongjiang. Echinococcosis is an important parasitic disease that harms humans and animals in northern and southwestern my country and is a statutory infectious disease. This standard was proposed by the Ministry of Health of the People's Republic of China. The responsible drafting unit of this standard: Training Base for Echinococcosis Prevention and Control of the Ministry of Health (Xinjiang Institute of Endemic Disease Prevention and Control); participating drafting units: People's Hospital of Xinjiang Uygur Autonomous Region, Institute of Parasitic Diseases, Chinese Academy of Preventive Medicine. The main drafters of this standard: Chai Junjie, Tan Jiazhong, Xue Haichou. This standard is interpreted by the Chinese Academy of Preventive Medicine, the technical unit entrusted by the Ministry of Health of the People's Republic of China.
1 Scope
This standard specifies the diagnosis and treatment principles of cystic echinococcosis and alveolar echinococcosis. It is applicable to the diagnosis and treatment of both types of echinococcosis by medical care, health and epidemic prevention and endemic disease prevention and control institutions at all levels and types.
2 Definition
2.1 Cystic echinococcosis cystichydatiddisease (CHD), cysticechinococcosis (CE)
Disease caused by the parasitism of the larvae of Echinococcus granulosus.
2.2 Alveolar echinococcosis alveolarhydatiddisease (AHD), alveolarechinococcosis (AE)
Disease caused by the parasitism of the larvae of Echinococcus multilocularis.
3 Diagnostic criteria
3.1 Epidemiological history
Residence or travel history in endemic areas, raising or contacting domestic dogs. History of field work and fox hunting; contact with fox bodies, skins, etc. 3.2 Clinical manifestations
3.2.1 Cystic echinococcosis: There may be no symptoms in the early stage, and it is often found in imaging examination. Hepatic cystic echinococcosis has dull pain in the liver area, upper abdominal fullness, indigestion, weight loss, and anemia. Enlarged liver and upper abdominal mass. Pulmonary cystic echinococcosis has dull pain, tingling, chest tightness, cough, shortness of breath, hemoptysis, and sometimes powdery internal capsule fragments or daughter cysts are coughed up with sputum, or the head hook of the protoscolex is found in sputum examination. Other organ echinococcosis has the unique symptoms of the space-occupying disease of the organ.
3.2.2 Alveolar echinococcosis: enlarged liver, dull pain in the liver area. In the late stage, liver function is impaired, spleen is enlarged, hard nodules and jaundice can be touched in the liver, and weight loss and exhaustion occur. When metastasis occurs, symptoms occur in the organs where the lesions are located.
3.3 Imaging examination
Imaging features are of great value in the diagnosis of echinococcosis. B-ultrasound is the main method for hepatic echinococcosis, and X-ray examination is the main method for pulmonary echinococcosis. Each has characteristic images. For differential diagnosis or when there is a special need, CT examination can be performed (see Appendix B for details). 3.4 Laboratory examination (see Appendix A for details)
3.4.1 Detection of specific antibodies in serum (using IHA, ELISA, EITB, etc.).
3.4.2 Detection of specific circulating antigens or immune complexes in serum. 3.4.3 Parasitological examination of sputum and expectorated material. 3.4.4 Pathological histological examination of clinical specimens. 3.5 Clinical diagnosis
Those with epidemiological history, major clinical symptoms or signs, imaging features or specific antibodies detected in serum.
3.6 Confirmed diagnosiswwW.bzxz.Net
Those who meet one of the following conditions in addition to the basis for clinical diagnosis. 3.6.1 Specific circulating antigens or immune complexes are repeatedly detected in serum 3.6.2 Head hooks are detected in coughed-up capsules, asci or sputum. 3.6.3 Confirmed by pathological histological examination of clinical biopsy materials. 3.6.4 Surgical exploration confirms that it is a hydatid cyst. Treatment principles
Surgical treatment
Surgical methods for cystic echinococcosis
4. 1. 1.1
Puncture and removal of the internal capsule.
4. 1. 1. 2
Complete removal of the internal capsule.
4.1.1.3 Complete removal of the hydatid (liver lobectomy). 4.1.1.4)
Reasonable treatment of residual cysts outside the liver and lungs.
4.1-2 Surgical methods for alveolar echinococcosis
4. 1. 2. 1
Radical resection.
4. 1. 2. 2
Palliative resection.
Palliative drainage.
4.2 Drug treatment
4.2.1 Drugs and courses of treatment
Albendazole is the drug of choice, and a low-dose (10 mg/kg) long-term or multiple courses of repeated treatment is better. Patients with alveolar echinococcosis should be treated with drugs as the main treatment. For patients in the advanced stage, the course of treatment can be up to 3 to 5 years, or even lifelong maintenance treatment. 4.2.2 Indications
4.2.2.1 Cystic echinococcosis.
4.2.2.1.1 Secondary abdominal and thoracic echinococcosis. 4.2.2.1.2 Patients with echinococcosis in multiple organs, multiple cysts or recurrence after multiple surgeries. 4.2.2..1.3 Patients with echinococcosis discovered at an early stage. 4.2.2.1.4 Patients who are not suitable for surgery or refuse surgery. 4.2.2.2 Alveolar echinococcosis: Regardless of surgery or not, drug treatment should be carried out, including advanced patients and patients with lung and brain metastasis.
4.2.2.3 Patients undergoing 2 to 3 courses of treatment before and after surgery have the effect of controlling the transplantation of protoscoleces during surgery and preventing recurrence.
4.2.3 Contraindications
Forbidden to use in patients with impaired liver and kidney function and pregnant women.
5 Prevention
5.1 Implement dog management and praziquantel deworming. 5.2 Carry out health education on the prevention of echinococcosis. 5.3 Strictly implement slaughter management and properly handle the organs of infected livestock. 5.4 In areas with high incidence of alveolar echinococcosis, take measures to control the spread of the disease by foxes. 5.5 Conduct a census among the population in high-prevalence areas, find patients early and give them treatment. Appendix A
(Standard Appendix)
Immunological diagnostic methods
A1 Indirect hemocyte agglutination test (IHA)
A1.1 Antigen preparation
Sheep red blood cells are aldehyded with 2.5% glutaraldehyde, treated with echinoic acid at a final concentration of 1:20000, and sensitized with the optimal concentration of cyst fluid crude antigen or purified antigen. The sensitized red blood cells are made into a 5% suspension in pH7.2 PBS containing 10% sucrose and 1% normal immune serum, and are divided into aliquots (1mL/tube), stored in a refrigerator at 4℃ or freeze-dried and sealed (0.2mL/tube). The concentration used is 1%.
A1.2 Operation method
A1.2.1 Prepare antigen: If it is a freeze-dried product, add 1mL of distilled water to dilute and mix well for use.
A1.2.2 Use a micropipette to add 3 drops of 1% rabbit serum-phosphate buffered saline to the first well of the V-shaped microplate, and 1 drop (25μL) to each of the remaining wells; at least six wells should be made. Add 1 drop (25μL) of the serum to be tested to the first well, that is, the serum dilution ratio is 1:4; after mixing, draw 1 drop of serum from it and add it to the second well; mix well, and make multiple dilutions in turn; after mixing the last well, draw 1 drop and discard. And do negative and positive controls in parallel. A1.2.3 From the second well onwards, add 1 drop (25μL) of sensitized red blood cell suspension to each well; shake immediately for 1min; cover the plate, let it stand at room temperature for 1h, and observe the results. A1.3 Result judgment
A1.3.1 Negative reaction: All red blood cells sink to the bottom of the well in the form of dots, with smooth edges. A1.3.2 Positive reaction: Those with a positive reaction (++ or +++) when the serum is diluted 1:128 are judged as positive for hydatid serum antibodies. +++ Red blood cells form a thin layer of agglutination, filling the entire bottom of the well or with wrinkles on the edge ++ Red blood cells form a thin layer of agglutination, with a small area and loose edges. + Most of the red blood cells sink to the bottom of the well, forming a dot or circle, with blurred periphery or a small white dot in the center.
A2 Enzyme-linked immunosorbent assay (ELISA)
A2.1 Antigen
Antigen purified from hydatid cyst fluid (prepared by phosphotungstic acid and magnesium chloride precipitation method). A2.2 Operation method
A2.2.1 Antigen coated polystyrene plate: Dilute the antigen to the optimal concentration with 0.05mol/L pH9.6 carbonate buffer, add 100μL to each concave well, and place in a temperature box. Incubate at 4℃ overnight (or 12-24h). The next day, remove the antigen and wash three times with phosphate buffered saline (0.01mol/L, pH7.4PBS-T) containing 0.05% Tween 20, 5min each time, and spin dry. A2.2.2. Add the serum to be tested: dilute the serum with PBST at 1:200, add 100μL to each well, and set one positive reference well, three negative reference wells and one PBS control well for each plate. Place in a temperature box at 37℃ for 1h, then take out, pour off the serum, and wash as before. A2.2.3 Add conjugate: add 100μL of horseradish peroxidase (HRP)-labeled conjugate with PBS-T as working dilution, 37℃, 1h, pour off the conjugate, and wash as before.
A2.2.4 Add substrate: usually add 100μL of o-phenylenediamine (OPD) substrate solution (10mgOPD+25mLpH5.0 citric acid buffer+30%H20210μL), 37℃, 30min. A2.2.5 Add stop solution: 2mol/LH2SO450μL. A2.3 Result judgment
Read the 492nmOD value on the enzyme-labeled colorimeter, and the S/N value of the sample to be tested (S) to the negative control serum (N) ≥2.1 is the positive critical valueA3PVC film rapid ELISA
A3.1 Antigen
Purification of antigen from hydatid cyst fluid.
A3.2 Operation method
A3.2.1 Take the PVC film soft plate coated with antigen, number it, wash it once with PBS-T, and then add 200μuL of PBS-T to each well; A3.2.2 Add 10μL of the serum to be tested and the reference serum (one well for negative control and one well for positive control on each plate) to each concave well, mix well, and place in a humidified box at 37℃ for 5min (or 25℃ for 10min). Pour off the serum, wash it 8 times in a row with PBS-T, and spin dry. A3.2.3 Add enzyme conjugate: dilute according to working concentration, 200μL per well, 37℃5min, pour off the conjugate, wash 8 times as above, wash once with distilled water, shake ten. A3.2.4 Add substrate solution: TMB substrate containing 3% H2O2, 200μL per well, react for 5-10min (Preparation of TMB substrate solution: TMB 50mg is dissolved in 10mL dimethyl sulfoxide as mother solution, stored at 4℃. Take 1mL of mother solution + 50mL of pH 5.0 citric acid buffer + 30% H2O2 8μL before use).
A3.3 Result judgment
Judge the result by yourself according to the negative and positive controls of each batch. Negative is basically colorless, and positive is bright blue.
A4 Enzyme-linked immunosorbent technology (EITB)
A4.1 Preparation of antigen membrane strips
Use 5%~20% thick 0.75mm gradient gel, add 1ug/mm of hydatid cyst fluid antigen, the upper tank buffer is 0.1mol/L boric acid buffer, the lower tank liquid is 0.424mol/LpH9.18Tris/HCl, the antigen is separated by SDS-PAGE electrophoresis, and then transferred by electrophoresis. The transfer electrophoresis buffer is 0.212mol/LpH9.18Tris/HCL, containing 20% ​​methanol for 30min, that is, the separated antigen protein band is transferred from the gel to the nitrocellulose membrane, the membrane is washed 3 times with PBS-T, and 1 time with PBS, each time for 5min, and then the membrane is cut into 3mm wide strips, which are antigen membrane strips, sandwiched in PBS wet filter paper, sealed in a plastic bag, and stored in the refrigerator for use. If stored in a low-temperature refrigerator, it can be stored for a long time. A4.2 Operation method
A4.2.1 During the test, take out or thaw the membrane. First, add 495μL of PBS containing 5% skim milk powder and 3% Tween to the reaction tank, then add 5uL of serum to be tested, and finally add the antigen membrane strip. Each batch of tests should have a reference positive, negative and PBS control. Shake at room temperature for 1 hour (4℃ can be overnight). The next day, wash 4 times with 0.3% Tween-PBS, each time for 5 minutes and dry. A4.2.2 Add the conjugate diluted at the working concentration, room temperature for 1 hour, and wash as before. A4.2.3 Add DAB-H2O2 system (25mg3,3'-diaminobenzidine + 50mL Tween-free PBS + 5μL H2O2) to color for 10 minutes, rinse with tap water to stop the reaction, and dry.
A4.2.4 Interpretation of results: Visually inspect specific bands, with 60KD, 36KD, 32KD, 24KD, 20KD, 17KD and 12KD as positive bands. Among them, as long as the 12KD, 17KD24KD bands appear, they can be judged as positive.
A5 Circulating antigen detection
A5.1 Dilute the anti-hydatid monoclonal antibody to the working concentration with 0.05mol/LpH9.6 carbonate buffer to coat the ELISA plate, 100uL per well, wash the plate 3 times with PBS-T after overnight at 4℃.
A5.2 Add 100μL of PBS-T containing 10% calf serum to each well, seal the plate at 37℃ for 1h, and wash the plate 3 times.
A5.3 Dilute the serum to be tested with the above solution to 1:2, add 100uL to each well, add two wells for each specimen, incubate at 37C for 1h, and wash the plate 3 times. A5.4 Add 100μL of HRP-labeled monoclonal antibody at working concentration and incubate at 37℃ for 1h, then wash the plate 3 times.
A5.5 Add 100μL of OPD substrate solution and incubate at 37℃ for 30min. A5.6 Add 50oμL ​​of 2mol/LH2SO4 to terminate the reaction. A5.7 Measure the OD490 of each well.
A5.8 Control setting and result judgment: Set one positive control well (5-10μg/mL antigen 100uL) and three negative serum control wells on each plate, and take the negative control OD mean plus 3SD as the positive critical value.
A6 Circulating immune complex detection
A6.1 Use 0.01mol/L pH8.4 borate buffer to prepare PEG6000 into a 7% solution. Take 100μL of this solution and add an equal amount of serum to be tested in a ratio of 1:2. Mix at room temperature for 1 hour and centrifuge at 3000r/min for 1 hour.
A6.2 Aspirate the supernatant and add 0.5mL of 0.05mol/L pH9.6 carbonate buffer containing 8mol/L urea to the precipitate to dissolve the precipitate. A6.3 Use this solution to coat each well of the ELISA plate with 100μL. Coat two wells for each specimen. After overnight at 4℃, wash the plate three times (positive and negative controls are treated in the same way). A6.4 Add 100μL of PBS-T containing 10% calf serum to each well, seal the plate at 37℃ for 1 hour, and wash the plate three times.
Add 100μL of HRP-labeled anti-echinococcosis monoclonal antibody at working concentration, incubate at 37℃ for 1h, then wash the plate 3 times.
A6.6Add 100μL of OPD substrate, incubate at 37℃ for 30min. A6.7
Add 50uL of 2mol/LHSO4 to terminate the reaction. Measure OD4904 of each well
Control setting and result judgment: Set one positive control well (same as item 6.8) and three negative serum control wells on each plate, and take the mean OD value of negative control plus 3SD as the positive critical value. A7 Alveolar echinococcosis specific EITB technology
A7.1 Preparation of antigen membrane strips and immunoblotting test A7.1.1 Collect the cysts in the abdominal cavity of the long-clawed sand rat artificially infected with Echinococcus multilocularis, cut them into pieces and collect the protoscoleces, wash them repeatedly, dissolve them with sodium deoxycholate solution and extract them, dialyze them with PBS, collect the antigens, determine the protein content and adjust it to 2mg/mL concentration. A7.1.2 Perform SDSPAGE electrophoresis as usual using 12% or 20% polyacrylamide gel or 8%~16% gradient gel (thickness 1.5mm). Add 400ug of protein to each large hole on the gel with a length of 6cm. After electrophoresis, transfer to nitrocellulose membrane (pore size 0.45um) as usual. Then cut into membrane strips and perform immunoblotting test. The serum to be tested is diluted 1:50. A7.2 Interpretation of results: The enzyme immunostaining band at the molecular weight of 18KD is positive.
Appendix B
(Standard Appendix)
Basic features of imaging diagnosis of echinococcosis
B1Basic features of ultrasound images
B1:1 Hepatic cystic echinococcosis: Simple cysts appear as dark areas with clear boundaries and no echoes in the liquid, with enhanced echoes on the posterior wall. Large and complete cysts can be seen with double walls. Mature cysts show floating light spots inside the cysts and light spots deposited at the bottom due to the increase in cystic sand. When the internal capsule is separated, it presents a typical double-wall structure, which has specific diagnostic significance. When the internal capsule ruptures, the "water lily sign" can be seen in the cyst fluid. Cysts with multiple daughters show unevenly thick and thin high-echo partitions inside the cysts, forming peak-shaped or wheel-shaped structures. Solidified cysts appear as high-echo solid masses without enhanced shadows on the posterior wall, which are difficult to distinguish from tumors. The calcified cyst wall is highly echogenic, accompanied by acoustic shadows, and the shape can be ring-toothed, dot-shaped, or a large number of small rings. B1.2 Alveolar echinococcosis: It shows abnormal echo areas of solid space-occupying lesions in the liver, which appear as dense strong light spots or light clusters. When there is central necrosis, it shows liquid dark areas. B2 Basic characteristics of x-ray images
B2.1 Pulmonary cystic echinococcosis: Pulmonary cysts with a diameter of less than 2 cm are spherical shadows with low density, rough edges, and unclear. Larger cysts have clear outlines, neat edges, sharp boundaries, and uniform density. They are round, oval, or lobed with notches, and are single or multiple isolated solid shadows. Due to the compression of the cysts, the trachea and heart may be displaced. The cysts in the lower lobes of the lungs may show the characteristics of deformation with breathing (echinococcosis respiratory disease). B2.2 Hepatic cystic echinococcosis: Abdominal plain films show that the liver contour is enlarged, and the right diaphragm of the cyst at the top of the liver bulges or protrudes into the chest cavity. Larger cysts can raise the diaphragm, weaken the respiratory movement, and even squeeze the heart to the left. The cysts in the middle and lower parts of the liver do not raise the diaphragm significantly, and a hemispherical shadow with a higher density can be seen at the lower edge of the liver. Calcification shadows can be seen when the cysts are calcified. B3 Basic characteristics of CT images
B3.1 Hepatic cystic echinococcosis
The liver contour is enlarged, and spherical space-occupying shadows of varying sizes are shown in the liver parenchyma. The inner cyst wall is smooth, 1-3 mm thick, and has a CT value of 10-20 Hu. The cyst is filled with fluid and has a watery density, with a CT value of <10 Hu. The outer cyst wall is thicker, 3-8 mm, and can show a double wall sign, with a CT value of 30-
Tip: This standard content only shows part of the intercepted content of the complete standard. If you need the complete standard, please go to the top to download the complete standard document for free.