Some standard content:
GB15991-1995
According to my country's "Law on the Prevention and Control of Infectious Diseases", plague is the first highly contagious disease, and it is also a natural epidemic disease. According to experts' estimates, the distribution area of plague's natural epidemic sources in the world has not decreased since the third world pandemic. Because its epidemic has a certain interval, people often have an illusion that plague is an ancient disease and will no longer pose a huge threat to humans. However, this is not the case. Over the years, countries with plague epidemic sources in the world (including my country, especially Yunnan) have occasionally experienced local outbreaks. The 1994 Indian plague pandemic reminded people that if the monitoring and control of plague is neglected, it can still cause great danger to humans.
At present, whether plague can cause a pandemic in the human world depends first on the accurate diagnosis of the first case. Only early diagnosis and early control can prevent it from becoming popular in the human world. For this reason, my country is in urgent need of a scientific, accurate, state-issued, and legally binding "Diagnostic Criteria for Plague".
During the drafting process, this standard was mainly based on the "Law of the People's Republic of China on the Prevention and Control of Infectious Diseases", and also referred to the "Plague Prevention Manual" of the former Soviet Ministry of Health, the WHO "Plague Manual" and the "Diagnosis of Plague" of the United States. The main reference materials are the relevant regulations that have been effective in my country for many years.
During the drafting process, this standard has been discussed three times by the Plague Expert Advisory Group of the Ministry of Health, and written reviews by relevant experts across the country. Finally, it was reviewed and approved by all members of the Subcommittee on Disinfection Standards for Infectious Diseases of the Ministry of Health. Appendix A, Appendix B, and Appendix C of this standard are all appendices to the standard. This standard was proposed by the Ministry of Health of the People's Republic of China. The drafting units of this standard are: Institute of Epidemiology and Microbiology, Chinese Academy of Preventive Medicine, Institute of Endemic Disease Control and Prevention, Qinghai Province, Institute of Endemic Disease Control and Prevention, Gansu Province, Institute of Plague Control and Prevention, Guangdong Zhanjiang, and Institute of Epidemiology and Prevention, Yunnan Province. The main drafters of this standard are: Liu Yunpeng, Zhu Jinqin, Wang Wenshao, Yu Dongzheng, Zeng Biaocheng, and Huang Jianhua. This standard is interpreted by the Office of Supervision and Management of Infectious Disease Control and Prevention of the Ministry of Health, the technical unit entrusted by the Ministry of Health. 340
National Standard of the People's Republic of China
Diagnostic criteria of plague
Diagnostic criteria of plagueGB15991—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 my country on the Prevention and Control of Infectious Diseases. 1 Scope
This standard specifies the criteria for determining the diagnosis of plague cases infected by hosts, vectors and other infected animals and animal products in seven types of epidemic sources in China, or infected by contact with plague laboratories and their experimental supplies, as well as quarantined cases entering my country from foreign plague-endemic areas.
This standard is applicable to plague monitoring and the determination of plague animal diseases. 2 References
The provisions contained in the following standards constitute the provisions of this standard through reference in this standard. When this standard is published, the versions shown are valid. All standards are subject to revision, and parties using this standard should explore the possibility of using the latest version of the following standards. GB15978-1995 Standards and principles for handling human plague epidemic areas GB15992-1995 Principles and methods for plague control and its assessment 3 Diagnostic principles
3.1 Patients have epidemiological clues.
3.2 In addition to clinical symptoms of plague and 3.1, patients must have a positive result of plague bacteriological diagnosis or passive hemagglutination test (PHA) serum F1 antibody diagnosis to be diagnosed.
4 Diagnostic standards
4.1 Epidemiological clues
10 days before the onset of the disease, the patient has been to the plague animal disease epidemic area or has been in contact with the source animals, animal products and plague patients in the plague epidemic area, entered the plague laboratory or been in contact with plague laboratory supplies. 4.2 Sudden onset, high fever, dramatic increase in white blood cells, without the use of antibacterial drugs (penicillin is ineffective), the condition rapidly deteriorates within 24 hours and has one of the following syndromes:
4.2.1 Acute lymphadenitis, swelling, severe pain and forced posture. 4.2.2 Severe toxemia and shock syndrome without obvious lymph node swelling. 4.2.3 Cough, chest pain, bloody sputum or hemoptysis. 4.2.4 Severe conjunctivitis with severe edema of the upper and lower eyelids. 4.2.5 Bloody diarrhea with severe abdominal pain, high fever and shock syndrome. 4.2.6 Painful red papules appear on the skin, which gradually bulge and form bloody blisters with gray-black surroundings and hard base. When the blisters rupture, the wound surface also turns gray-black.
4.2.7 Severe headache, lethargy, stiff neck, delirium, high brain pressure, turbid cerebrospinal fluid Approved by the State Administration of Technical Supervision on December 15, 1995, and implemented on July 1, 1996
GB15991-1995
4.3 Lymph node puncture fluid, blood, sputum, pharyngeal and eye secretions, and specimens from body organs or tubular bones of patients, isolated from plague bacteria.
4.4 Serum was collected from patients twice (10 days apart), and the F1 antibody detected by PHA method showed a 4-fold increase or more. 5 Suspected cases
Meet any one of 4.1 plus 4.2.
6 Confirmed cases
Suspected cases plus 4.3 or 4.4.
7 Latently infected persons
Those with epidemiological clues of plague, but no obvious clinical manifestations of plague, who have not been vaccinated with plague vaccine, and whose serum has an F1 antibody titer of 1:40 or above in PHA test.
8 Retrospective diagnosis cases
Those with epidemiological clues of plague, who have had clinical manifestations of plague, who have not been vaccinated with plague vaccine, and whose serum has an F1 antibody titer of 1:40 or above in PHA test. 9 Disease type
9.1 Confirmed cases of plague with clinical manifestations of 4.2.1 are bubonic plague. 9.2 Confirmed cases of plague with clinical manifestations of 4.2.2 are septicemic plague. 9.3 Confirmed cases of plague with clinical manifestations of 4.2.3 are pneumonic plague. 9.4 Confirmed cases of plague with clinical manifestations of 4.2.4 are ocular plague. 9.5 Confirmed cases of plague with clinical manifestations in 4.2.5 are considered enteric plague. 9.6 Confirmed cases of plague with clinical manifestations in 4.2.6 are considered cutaneous plague. 9.7 Confirmed cases of plague with clinical manifestations in 4.2.7 are considered meningeal plague. 3.12
A1 Sampling of suspected plague patients
GB15991-1995
Appendix A
(Standard Appendix)
Taking, storing and transporting suspected plague materialsA1.1 Before taking antimicrobial drugs, samples should be taken from suspected plague patients according to their symptoms and signs at the following specified sites. A1.2 In addition to taking samples from the corresponding sites, 3-5 mL of venous blood should be taken from patients of all types of suspected plague for bacterial testing and serological diagnosis.
A1.3 Sampling of patients suspected of bubonic plague
A1.3.1 Select enlarged lymph nodes, disinfect them locally with iodine and alcohol, fix them with the thumb and index finger of the left hand, pierce the lymph nodes with a sterile syringe (12-16 gauge needle), extract an appropriate amount of tissue fluid, store it in a sterile test tube or directly inoculate it on a blood agar plate. A1.3.2 For patients with less obvious enlargement of lymph nodes, 0.3-0.5 mL of sterile saline can be injected into the lymph nodes first, and then the fluid can be extracted after a short pause. A1.3.3 In the late stage of infection, tissue fluid can be extracted by puncturing around the enlarged lymph nodes. A1.4 Sampling of patients suspected of pneumonic plague
A1.4.1 Ask the patient to cough on the blood agar plate, or collect the bloody sputum specimen in a sterile plate III or wide-mouth bottle for examination. A1.4.2 Use a sterile cotton swab to smear the pharyngeal secretions, and store the swab in a sterile test tube or sterile saline tube for examination. A1.5 For suspected septicemic plague, more than 1 mL of venous blood should be collected. A1.6 For suspected ocular plague, use a cotton swab or a sterile capillary pipette to collect ocular secretions. A1.7 For suspected intestinal plague, feces should be collected for examination. A1.8 Sampling for suspected cutaneous plague
A1.8.1 During the vesicular and pustular stages, the surface of the pustule can be disinfected with alcohol, and a sterile syringe can be inserted into the blister from the side to extract the contents for examination.
A1.8.2 During the ulcer and induration stages, use sterile tweezers to hold a sterile cotton ball to smear the ulcer surface and the wound surface under the skin, and store the cotton ball in a sterile test tube or sterile saline for examination.
A7.9 For patients with suspected meningitis plague, cerebrospinal fluid can be extracted by lumbar puncture for examination. A1.10 For close contacts of plague patients, contacts of plague-contaminated materials, and suspected plague patients who do not show typical suspicious symptoms in the early stage, samples should be collected for examination according to the provisions of A1.2 and A1.4. A2 Collection of samples from suspected plague bodies
A2.1 The first suspected plague body should be dissected. A2.1.1 Before collecting samples, preparations should be made for dissection equipment, site selection, and body handling. A2.1.2. Take the liver, spleen, lung, heart blood and lymph nodes with suspected pathological changes in a sterile manner and store them in sterile flat dishes or test tubes. When the body shows signs of decay, long bone materials must be taken. A2.2. If dissection is not possible, local sampling can be performed. Use a lumbar puncture device to puncture and take tissues in the order of lymph nodes, heart, liver, spleen and lungs, and store them in sterile test tubes. When the body is decayed, bone marrow can be punctured. A3. All materials taken should be stored in sterilizer III; tissue blocks can be stored in sterile saline or 5~~10mL Broke's solution; Cary Bleir culture medium can also be used to store and transport materials. The container is sealed with paraffin. A4. The test materials should be tightly packaged, stored in a suitable place, and stored at a temperature not higher than 4C. A5. Fill in the inspection form accurately and in detail.
A6. Appoint two personnel (including professionals) to deliver the test materials by fast transportation. Deliver directly to the professional laboratory responsible for inspection work in the area.
GB15991-1995
A7 When receiving materials, first check the packaging. There must be no damage or contamination. If there is any damage, disinfect it immediately and report it to the relevant unit for processing. When receiving materials, check the type and quantity according to the list and record the signature accurately. Appendix B
(Standardized Appendix]
Plague bacteria inspection procedures and result judgment
B1 Basic requirements for plague bacteria inspection
B1.1 Plague bacteria inspection must be carried out in a dedicated laboratory. B1.2 Inspectors must be familiar with the laboratory management system, self-protection system, technical operation procedures, etc. B1.3 Anyone entering the germ room must work with more than two people at the same time. B1.4 Make timely and accurate records of various laboratory tests. B2 Reverse hemagglutination test
B2.1 For samples from suspected plague patients and suspected plague households, as long as the sample volume allows, reverse hemagglutination tests should be carried out to make early diagnosis.
B2. 2 The operation steps and result judgment of the reverse hemagglutination test are shown in Appendix CB2.3 If a positive result is found with a reverse hemagglutination titer of 1100 or more, the unit responsible for the inspection shall make a suspected plague report within 24 hours after receiving the sample.
B2.4 Reverse hemagglutination test specimens and various equipment used should be regarded as contaminated items and must be soaked in 5% Lysol for 24 hours before washing.
B3 Bacterial culture
B3.1 Fresh materials can be directly coated with hemolytic (0.1%) Herxheimer agar plates and streaked according to the three-stage method. B3.2 Corrupted materials It can be streaked on gentian violet (1:100,000-1:200,000) sodium sulfite (0.025%) plates or gentian violet hemolytic plates. B3.3 For liquid materials and bone marrow, use sterilized platinum ear to take specimens and streak them. The organ material should be first imprinted on the surface of the plate, then streaked with platinum ear. Cotton swabs can be directly applied to the surface of the culture medium. B3.4 Different materials from the same patient or household can be applied to the same plate surface in grids. Each material is inoculated into two plates, one for separation culture and the other for plague phage lysis test. B3.5 Culture in a 28C incubator and observe daily for 14 to 96 hours. To find colonies with typical morphology of plague bacteria. Plates without serious contamination must be cultured for 7α and can be discarded when no suspected plague colonies appear. B4 Plague phage lysis test
B4.1 On the plate used for phage lysis test in B3.4, drop a drop of phage on one side of the streak, and tilt the plate to make it flow vertically over the streak.
B4.2 When suspected plague colonies are found during isolation culture, use platinum ear to take the suspected colonies and re-streak them on the blood agar plate, and then add plague phage according to the above method.
B4.3 Place in a 28°C incubator for 24 hObserve for phagocytosis. Only when the phagocytosis bandwidth is wider than the traces of phage flow can the plague phage test be determined to be positive.
B5 Animal inoculation
B5.1 Patient and corpse materials, especially corrupt materials, must be inoculated with mice (18-20g) or guinea pigs (250344
~300g) at the same time as bacterial culture.
GB15991-1995
B5.2 Organ pieces are placed in a sterile mortar, cut into pieces with sterile scissors and grind into a homogenate, add an appropriate amount of physiological saline, and make a suspension for use. B5.3 Fresh materials can be inoculated intraperitoneally or subcutaneously, guinea pigs are inoculated with 0.5-1.0mL, and mice are inoculated with 0.2-0.4mL. B5.4 Corrupted materials can be inoculated percutaneously. Shave the animal's abdominal hair and create slight scratches. Apply the material to the shaved skin with a cotton swab and rub it. Cover it with a flat dish cover during rubbing to prevent the material from splashing. B5.5 After inoculating the experimental animals, mark them and put them in a cage. Hang a tag to record the number, inoculation date, route, etc. Feed them 1 to 2 times a day until the animals die or are killed for autopsy after 7 days. B5.6 After the animals die, test them according to B3 and B4. B5.7 If the animals do not die after 7 days, they should be killed, and the spleen and suspected lesions should be made into homogenate and inoculated into the second group of animals. The animals should be killed after death or observation for 7 days, and tested according to B5.6. At the same time, serum should be collected for passive hemagglutination test. A negative report can be made only if there is no positive finding.
B6 Basis for determination of plague bacteria
B6.1 Plague colonies are isolated and the plague phage lysis test is positive according to the standard of B4.3, and then the bacteriological determination of plague can be made. The unit responsible for the inspection shall submit the plague bacteria identification report to the responsible department within 24 hours after the phage lysis test results are produced. And make a confirmation or correction report on the suspected plague report made in the past. B6.2 If the experimental animal dies, the plague bacteria are re-isolated from the dead experimental animal and confirmed by the phage lysis test, the unit responsible for the inspection shall make an identification report of the highly virulent plague bacteria. Appendix C
(Standard Appendix)
Operational steps and judgment of indirect hemagglutination test and reverse hemagglutination test
C1 Indirect hemagglutination test (PHA)
C1.1 During the acute and recovery period of suspected plague patients, as well as people with a suspected history of plague, venous blood should be taken for indirect hemagglutination test to check whether there are anti-plague F1 antibodies in the blood. C1.2 Test materials
C1.2. 1 35mL of venous blood should be taken from patients with various types of plague to separate serum. C1.2.2 The serum to be tested should be inactivated for 30 minutes for diagnosis. C1.3 Reagents
C1.3.1 2.5% F1 antigen sensitized blood cells.
a) Use plague whole bacteria immune serum with a bacterial agglutination titer of 1:320, and the hemagglutination titer should not be less than 1:40000. b) The cross titer with pseudotuberculosis serum should not be higher than 1:5. Sensitized blood cells that meet a) and b) are qualified diagnostic blood cells. The above titration should be performed before the start of each plague season or when the sensitized blood cell batch number is changed. bzxZ.net
C1.3.21% normal rabbit serum saline.
C1.3.32.5% tannic acid blood cells
Cl.3.4 F1 antigen solution (50~100 μg/mL). C1.4 Operation steps
C7.4.7 Take two rows of small test tubes (15mm×100mm), add 0.9ml of 1% normal rabbit serum saline to the first tube in the first row, and add 0.5ml to each of the remaining tubes345
GB15991-1995
, which is the hemagglutination inhibition column, and add 0.25ml to each tube in the second row, which is the hemagglutination column. C1.4.2 Take 0.1mL of the test serum and add it to the first tube in the first column. Perform double-row parallel dilution as shown in Table C1. After mixing the first tube, pipette 0.75mL, of which 0.25mL is added to the first tube in the second column, and 0.5mL is added to the second tube in the first column. After mixing, pipette another 0.75mL, and dilute as before until the last tube, discarding 0.5mL. Table C1 Test tube hemagglutination and hemagglutination inhibition test operation (two-row test) Test materials
First column Hemagglutination inhibition
1% normal rabbit serum saline
Test serum
Second column Hemagglutination
1% normal rabbit serum saline
1:40
1:320
1:1280
1:2560
-0.50.5 Discard
C1.4.3 Add 0.25mL F1 antigen solution (50-100μg/mL) to each tube in the first column, mix well and place in a 37℃ incubator for 10-15min. C1.4.4 Add one drop (0.05mL) of 2.5% F1 antigen-sensitized blood cells to each tube, mix well and place in a 37℃ incubator, and observe the results after 2h. C1.4.5 The following control tubes are set up at the same time as the test: a) 0.5mL of 1% normal rabbit serum saline + 0.05mL of 2.5% F1 antigen-sensitized blood cells; b) 0.5mL of 1% normal rabbit serum saline + 0.05mL of 2.5% tannic acid blood cells; c) 0.5mL of 1:20 test serum + 0.05mL of 2.5% tannic acid blood cells. C1.5 Result determination
C1.5.1 Each control tube should not show agglutination. C1.5.2 The second column is the test column, which is divided into the following types according to the degree of blood cell agglutination: a) "#" agglutinated blood cells spread all over the bottom of the tube, with obvious folded edges. When the antibody is excessive, the agglutination is loose and wreath-shaped; b) "++" agglutinated blood cells spread all over the bottom of the tube, without folded edges; c) "++" blood cells are not completely agglutinated, and the bottom of the tube is a neat circle, but there is very obvious blood cell agglutination inside and outside the circle; d) "+" the bottom of the tube forms a smaller circle, and there is only a small amount of blood cell agglutination inside and outside the circle. 346
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