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Delivery: <= 3 days. True-PDF full-copy in English will be manually translated and delivered via email. WS/T 191-2017: Diagnosis for chancroid Status: Valid WS/T 191: Historical versions
Basic dataStandard ID: WS/T 191-2017 (WS/T191-2017)Description (Translated English): Diagnosis for chancroid Sector / Industry: Health Industry Standard (Recommended) Classification of Chinese Standard: C59 Word Count Estimation: 14,168 Date of Issue: 2017-07-24 Date of Implementation: 2018-02-01 Older Standard (superseded by this standard): WS 191-1999 Regulation (derived from): State-Health-Communication (2017) 7 Issuing agency(ies): National Health and Family Planning Commission of the People's Republic of China WS/T 191-2017: Diagnosis for chancroid---This is a DRAFT version for illustration, not a final translation. Full copy of true-PDF in English version (including equations, symbols, images, flow-chart, tables, and figures etc.) will be manually/carefully translated upon your order.(Diagnosis of chancre) ICS 11.020 C 59 ws People's Republic of China health industry standards Replacing WS 191-1999 Softly under the diagnosis Diagnosis for chancroid 2017 - 07 - 24 Posted 2018 - 02 - 01 implementation People's Republic of China National Health and Family Planning Commission released ForewordThis standard was drafted in accordance with the rules given in GB/T 1.1-2009. This standard replaces WS 191-1999 "soft bottom 疳 diagnostic criteria and principles of treatment", this standard from the date of implementation, WS 191-1999 Abolished at the same time. This standard compared with WS 191-1999, the main changes are as follows. - The standard nature changed from mandatory to recommended. - Change the standard name to "Soft Subsidy Diagnosis"; - Added terms and definitions (see Chapter 2); - The treatment principle was deleted (see Chapter 3 of the.1999 edition); - Removed clinical cure (see Chapter 4 of the.1999 edition); - Removed management and prevention (see Chapter 5 of the.1999 edition); - Removed the fluorescent antibody staining method in Annex A (see A.5 of the.1999 edition); - Added Appendix A alkaline phosphatase test (see A.4.5); - Removed Appendix B soft-bottom 疳 treatment program (see the.1999 edition of Appendix B); - Added Appendix B Delivery and separation of Hautobacterium Duchenne commonly used medium (see Appendix B); - Added a description of Appendix C soft-ware ((see Appendix C). This standard was drafted. Chinese Academy of Dermatology Hospital (Institute), Jiangsu Provincial People's Hospital (Nanjing Medical University first attached Is a hospital), Shanghai Dermatology Hospital. The main drafters of this standard. Qi Shuzhen, Wang Chiaki, Su Xiaohong, Jiang Juan, Gong Xiangdong, Gong Kuanglong, Luo Dan, Zhou Pingyu. This standard replaces the standards previously issued as follows. - WS 191-1999. Softly under the diagnosis1 ScopeThis standard specifies the principles of soft under the diagnosis, diagnosis, diagnosis and differential diagnosis. This standard applies to all types of medical and health institutions at all levels and their medical staff on the soft under the diagnosis.2 Terms and definitionsThe following terms and definitions apply to this document. 2.1 Soft chancroid chancroid Painful and ulcerative sexually transmitted diseases caused by Duchenne's Haemophilus infection, often associated with inguinal lymphadenitis Lesions. 2.2 Haemophilus ducreyi Ducrey Haemophilus is a facultative anaerobes with no movement ability, no spores, negative Gram stain, short rod shape, blunt round ends, length of 1.6 μm ~ 2 μm, 0.5 μm wide, most of which are arranged in a chain or fish-like shape in the extracellular matrix, and a few are distributed in clusters in the cells.3 diagnostic principlesAccording to epidemiological history, clinical manifestations and laboratory tests for a comprehensive analysis to make a diagnosis.4 diagnostic basis4.1 Epidemiology history A history of unsafe sex, or a history of sexually transmitted infections, or multiple sexual partners. 4.2 clinical manifestations The incubation period of 3 d ~ 14 d, an average of 4 d ~ 7 d. Initial lesion for the genital area inflammatory papules, 1 d ~ 2 d quickly became pustules, 2 d ~ 3 d pustules ulceration pain Ulcer Ulcers were round or oval, the edge is not complete, the base covered with purulent secretions, remove the exudate, the basal vascular thick granulation Tissue, tenderness, hemorrhage; satellite ulcers may occur around the skin lesions. Ulcers in January to February healed, residual scar. Men occur in the coronary sulcus, foreskin, glans, penis dry, perineal and perianal, etc .; women occur in the size of the labia, urethra, vagina Mouth, vaginal wall, cervix, perineum and perianal and other places. Approximately 50% of patients develop acute painful purulent inguinal lymphadenitis within 1 week to 2 weeks after primary injury, Ditch and lymph nodes pain swelling, often unilateral, but also bilateral involvement. Swollen lymph nodes often fluctuate flu, can be natural ruptured pus, shaped Into a long and narrow shallow ulcer. 4.3 Laboratory tests (see Appendix A) 4.3.1 Microscopy Gram stains were taken from specimens of clinical ulcer secretions, and Gram stain negative for Brevibacterium was found (see Appendix A, A.2). 4.3.2 Cultivation Bacterial cultures of clinical ulcer secretions (see Appendix B for medium preparation), and biochemically identified Haemophilus ducreyi (see A.3 and A.4).5 diagnostic5.1 suspected cases Meet 4.2, with or without 4.1 and 4.3.1, and in line with. (1) ulcers occurred more than 7 d, ulceration with a dark field microscope Treponema pallidum syphilis can not be found, or syphilis serological test negative; (2) the clinical exclusion of herpes simplex virus (HSV) infection, HSV-PCR test or HSV antigen test positive. 5.2 confirmed cases At the same time in line with 4.3.2 and 5.1.6 differential diagnosisSoft bottom 一 should be with a syphilis hard 疳 疳, genital herpes and venereal lymphogranuloma differential diagnosis (see Appendix C C.5). AA Appendix A. (Normative) Laboratory tests for Haemophilus ducreyi A.1 specimen collection A.1.1 drawing tools Cotton swabs or calcium alginate swabs. A.1.2 drawn parts Genital ulcers or swollen lymph nodes. A.1.3 cutaneous mucosal lesions drawn Gently wipe the surface of the crust with a sterile cotton swab and dirt, and then take the secretion of swabs, drawing from the base of the ulcer or edge, Do smear or culture. A.1.4 Lymph node acquisition Select a fluctuating lymph nodes, disinfect the surface of the lymph nodes, dry with sterile dry cotton balls. With 1mL sterile syringe with 12 needle, suction Take normal saline 0.25 mL ~ 0.5 mL, sterilely punctured lymph nodes and saline, and then inhaled syringes, repeated 2 to 3 times, Take aspiration smear test or culture. A.2 Microscopic examination A.2.1 Materials Optical microscopes, sterile swabs, slides, syringes, and saline. A.2.2 Method Apply the specimen evenly and gently to a clean glass slide, then dry in air with flame fixed. A.2.3 Gram stain The steps are as follows. a) The fixed smear covered with crystal violet solution, dyed 30 s ~ 60 s, wash quickly in running water. b) covered with smear iodine, stained 30 s ~ 60 s, washed with running water. c) Decolourize with acetone-ethanol solution until the smear has no purple off. Usually 10 s ~ 20 s, depending on the thickness of the coating. avoid Free from decolorization, or gram-positive bacteria can be mistaken for negative bacteria. Rinse quickly in the water to stop decolorization, the excess water suction Water blot dry. d) counterstain with complex red liquid 1 min, rinsed with running water, blotted dry with absorbent paper. A.2.4 results and explanation Use 10 × 100 times oil mirror inspection. Haemophilus ducreyi is a Gram-negative, short rod-like, blunt-ended at both ends, 1.6 μm ~ 2 μm in length and 0.5 μm wide, mostly in cells Outside the chain or fish-shaped arrangement, a small number of intracellular distribution of clumps. Due to genital ulcers often have a variety of microbes living, direct smear clinical specimens Gram stain microscopy results are not reliable, false positive and false Negative, so smear check only for the initial judgment, not as a basis for the diagnosis. A.3 Haemophilus Duchenne transport and isolation and culture A.3.1 Materials A.3.1.1 Transport medium BM-SGA delivery medium. The main ingredients include. L-glutamine, calf albumin Ⅴ, 3 mg/L vancomycin. A.3.1.2 Isolate the medium A.3.1.2.1 Modified Thayer-Martin culture medium. The main ingredients include. GC basal medium powder, hemoglobin, 1% IsoVitaleX Increasing agent, 3 mg/L vancomycin. A.3.1.2.2 Chocolate agar medium. The main components include. meat extract (meat extract liquid) agar, aseptic defibrinated blood, 3 mg/L Elan Inoculation (medium preparation see Appendix B). A.3.2 culture conditions The specimens inoculated on the medium, zoned and separated. Media inoculated with specimens should be placed in relative humidity above 80%, culture temperature 33 ℃ ~ 35 ℃, 5% ~ 10% carbon dioxide environment (Candlestick) culture. 48 h ~ 72 h to watch the result. Plates that did not show positive results Should continue to observe the 7 d before discarding. A.3.3 initial identification of cultured colonies A.3.3.1 Colony characteristics. Haemophilus ducreyi after 48 h ~ 72 h culture, the diameter of 1 mm ~ 2 mm colonies can be formed, the colony is smooth, Was translucent, light gray or yellow-gray. Old culture, flat colonies, the color becomes gray to brown. With platinum ears can be complete The colonies along the agar surface to promote, that is, to promote positive test for Haemophilus Ducrei typical characteristics. A.3.3.2 Gram stain. take from the colonies smear, Gram stain (see A.2). A.4 biochemical identification test A.4.1 oxidase test A.4.1.1 Principle. Haemophilus Ducos imperfect enzyme system, but can produce weak oxidase, which produces oxygen ions can oxidase reagent (Dimethyl p-aniline hydrochloride) into quinone compounds, a weak color reaction. However, there are also reports of oxidase-negative strains. A.4.1.2 Method. The oxidase reagent is formulated into 0.5% ~ 1.0% aqueous solution. Drop the solution on suspect colonies and observe the color change. A.4.1.3 Results. After dropping 1% dimethyl-p-aniline hydrochloride solution, usually within 15 s ~ 20 s colonies that light pink, and then gradually Into deep purple, finally black. A.4.1.4 Clinical Significance. Oxidase test, cell morphology and colony morphology are three important criteria for the initial identification of Haemophilus ducreyi. A.4.2 catalase test A.4.2.1 Principle. Bacteria with catalase (catalase) catalyze the release of nascent oxygen by hydrogen peroxide and subsequent formation of oxygen molecules Now bubbles. A.4.2.2 Method (slide method). Pick the colonies on the medium, placed on a clean glass slide, and then add 1 drop of 3% to 6% hydrogen peroxide That is the observation. A.4.2.3 Results. In 30 s, the bubble generator is positive. A.4.2.4 Clinical Significance. Haemophilus ducreis does not produce air bubbles and therefore negative. A.4.3 porphyrin test A.4.3.1 Principle. Used to detect the ability of bacteria to convert δ-amino-γ-keto-valerate hydrochloride to porphyrin and porphobilinogen. Because Duke Haemophilus is a haemophilus that is strictly dependent on the growth of hemin. It does not convert δ-amino-γ-keto-valerate hydrochloride to porphyrin The test results for the negative reaction. A.4.3.2 Methods. Add 0.5 mL of 2 mmol/L hydrochloric acid-δ-aminolevulinic acid solution to a 12 mm × 75 mm test tube to prepare concentrated The bacterial suspension (1 × 109/mL) was placed in a 35 ℃ ~ 37 ℃ incubator for 4 h. Then test tube in the darkroom with wood lamp irradiation, observe the knot fruit. A.4.3.3 Results. The presence of red fluorescence indicates the presence of porphyrin is positive. A.4.3.4 Clinical Significance. Haemophilus ducreyi, which relies heavily on hemin, lacks the ability to convert to porphyrin and is therefore negative. A.4.4 nitrate reduction test A.4.4.1 Principle. The nitrate reduction reaction consists of two aspects. 1) bacteria in the process of anabolism, the nitrate is reduced to nitrite and nitrogen, and then by the ammonia into amino acids and other bacteria Nitrogen compounds. 2) During catabolism, nitrates or nitrites replace oxygen as the terminal hydrogen in the respiratory system. Nitrate reduction The process varies from bacterium to bacterium, Haemophilus ducreyi can reduce nitrate in the culture medium to nitrite, nitrite and reagent Acetic acid role, generating nitrous acid, nitrite and nitrobenzene sulfonic acid reagent in the role of diazo benzene sulfonic acid, and then with the reagent Of α-naphthylamine to form N-α-naphthylazobenzene sulfonic acid (red). A.4.4.2 Methods. Take 48 h culture made of bacteria suspension (109/mL), take 0.04 mL in a small test tube, add 0.04 mL of 0.05% NaNO3 solution and 0.04 mL of 25 mmol/L phosphate buffer (pH 6.8). Incubate 1 h at 35 ° C in a water bath. Add 0.5% p-aminobenzene separately Sulfonate acetic acid solution and 0.5% naphthylamine solution each 0.06 mL, shake, observe. A.4.4.3 Results. Within 2 min ~ 10 min color, pink positive, pink does not appear negative. A.4.4.4 Clinical significance. Ducrey Haemophilus nitrate reduction test was positive. A.4.5 alkaline phosphatase test A.4.5.1 Method. Prepare a suspension of concentrated bacteria (109/mL) in 0.5 mL of 0.03% sodium phosphate free disodium tube. Test tube 37 ℃ water bath After incubation for 4 h, add 4 drops of 0.5% 2,6-dibromoquinone-4-chloroimine in methanol, shake well and set at room temperature for 15 min. Add 0.3 mL n-butanol, shake Stir well for 5 min. Observed. A.4.5.2 Results. Blue but purple in the butanol layer is positive. A.4.5.3 Clinical significance. Duke Klenow Haemophilus alkaline phosphatase test was positive. A.4.6 Duke Haemophilus biochemical test results Hemophilus oxidase test positive, catalase test negative, porphyrin test negative, nitrate reduction test positive, alkaline phosphatase Enzyme test positive. A.4.7 Training significance and precautions Duke Klebsiella Haemophilus culture sensitivity of 60% to 80%, is currently recommended by the World Health Organization and the diagnosis of soft drinks Laboratory methods. Duke Haemophilus resistance to the environment is very weak, in order to improve the success rate of culture, specimens of the shorter the better, take Material should be immediately inoculated on the separation medium. Clinical specimens that can not be inoculated into the culture medium in time should be placed in the transport medium but must Transferred to the separation medium within a week. BBAppendix B(Informative) Preparation and delivery of Haemophilus ducreyi culture medium B.1 Delivery medium BM-SGA preparation B.1.1 BM composition BM composition is as follows. a) Disodium hydrogen phosphate (Na2HPO4) 10.0 g; b) Magnesium nitrate 0.1 g; c) potassium dihydrogen phosphate (KH2PO4) 2.0 g; d) sodium chloride 5.0 g; e) Calcium Chloride 0.1 g; f) Heme 0.2 g; g) Sodium thiglycollate 1.0 g; h) Bacto agar 7.5 g; i) 990 mL of distilled water; j) Adjust pH to 7.5. B.1.2 SGA composition SGA ingredients are as follows. a) selenium dioxide (SeO2) 0.003 mg/L; b) L-glutamine 3 mg/L; c) vancomycin 3 mg/L; d) bovine serum albumin (albumin) 2.0 g/L; e) Distilled water 10 mL. B.1.3 preparation method SGA components using a pore size of 0.45 μm membrane filter sterilization reserve. The BM ingredients were sterilized at 121 ° C for 15 min and allowed to cool to 50 ° C, sterile Add SGA solution to operate and dispense it in sterile test tube with screw cap, 6 mL for each tube, then put for 4 days at 30 ° C. B.2 separation medium preparation B.2.1 Preparation of modified Thayer-Martin medium B.2.1.1 Components B.2.1.1.1 GC basal medium powder Each 3.6 g GC basal medium powder can be equipped with 100 mL TM finished medium, which contains peptone 1.5 g, corn flour 0.1 g, hydrogen phosphate 0.4 g of dipotassium (K2HPO4), 0.1 g of potassium dihydrogen phosphate (KH2PO4), 0.5 g of sodium chloride (NaCl) and 1.2 g of agar. B.2.1.1.2 VCN bacteriostatic agent A solution of vancomycin (300 μg), colistin (750 μg) and nystatin 1250 units. For the inhibition of Proteus plus trimethoprim 500 μg. Use immediately after preparation or storage -20 ℃ below within 2 weeks Finish. B.2.1.1.3 IsoVitaleX enhancer The ingredients are added in 1 L of distilled water as follows. a) Vitamin B12 0.01 g; b) L-glutamine 10.0 g; c) P-aminobenzoic acid 0.012 g; d) adenine 1.0 g; e) guanine 0.03 g; f) diphosphopyridine nucleotide osidized (coenzyme I) 0.25 g; g) cocarboxylase 0.1 g; h) ferric nitrate 0.02 g; i) thiamine HCl 0.003 g; j) L-cysteine 25.9 g; k) L-cystine 1.1 g; l) dextrose 100 g. B.2.1.2 Preparation To 500 mL TM medium as an example, the specific preparation is as follows. a) Preparation of double-basal medium. Dissolve 18 g of GC base powder in 235 mL of distilled water (500 mL flask) Shake well, stirring while heating until boiling 1 min, so that it completely dissolved. b) Add 5 g of hemoglobin powder to 250 mL of water to make a 2% solution. When mixed, 5 g hemoglobin plus water 5 mL ~ 10 mL, Grind into a paste, and then gradually add distilled water, so that the entire solution into a slurry. Also available 2% hemoglobin solution finished. c) The above two liquids were 121 ℃ autoclaved 15 min and then cooled to about 50 ℃ standby. 2% hemoglobin solution is not finished Will be high pressure, just a little before the heat can be used. d) preparation of bacteria broth. add ampoules per ampoule attached to the attached with 10 mL, to dissolve. e) Preparation of bacteriostat. antibacterial agent per ampoule with aseptically add 5 mL of distilled water, shake, to fully dissolve. f) Aseptically mix 250 mL of 2% hemoglobin solution, 10 mL of broth, 5 mL of bacteriostat and 235 mL of GC medium. The total volume of 500 mL, can be inverted 70 mm plate 25 to 30. B.2.2 Preparation of chocolate agar B.2.2.1 Components Meat extract (or meat extract) agar (pH 7.2 ~ 7.4) 1 000 mL, sterile defibrinated blood (sheep or rabbit blood) 80 mL ~ 100 mL, Vancomycin 3 mg/L (or Polymyxin B 25 U/mL). B.2.2.2 Preparation The meat infusion agar autoclave, when cooled to 45 ° C add defibrinated blood (blood in the pre-37 ° C water bath preheated), shake Then placed in water bath, slowly heated to 85 ℃ ~ 90 ℃ for 5 min ~ 10 min, then cooled to about 50 ℃. To inhibit bacteria, Add vancomycin 3 mg/L (or polymyxin B 25 U/mL) when cooled to about 45 ° C. Pour gently on a sterile plate in. CCAppendix C(Informative) Soft under the brief introduction C.1 Etiology The carapace is a Duchenne Haemophilus, discovered by Ducrey in Italy in 1889. This bacteria facultative anaerobes, no exercise Ability, spore-free, Gram-negative, Brevibacterium, blunt round at both ends, 1.6 μm ~ 2 μm in length and 0.5 μm wide, mostly in the extracellular chain Or fish-like arrangement, a small number of intracellular distribution of clumps. C.2 Epidemiology Due to the lack of rapid and accurate laboratory testing of Haemophilus Duracheau, it is difficult to diagnose soft subduction and seriously affect soft 流 epidemiological assessment. According to the estimates of the World Health Organization, there are about 7 million cases of newly developed soft bottom per year in the world each year. In different regions and countries The prevalence of soft-shelled turtles is very different, the most common areas are Africa, the Caribbean, Southwest Asia or Southeast Asia. As in Kenya, Gangby Asia and Zimbabwe, the most common cause of genital ulcers is chancroid. There have been reports of sex workers in Kenya's capital Nairobi In the outbreak of soft bully. Anyone traveling to these areas who have unsafe sex may be able to bring the disease back to his home country. Sex workers in these areas If you work on sex work in Europe or elsewhere if you inflict soft tissue, you may also cause a small area of softness in Europe or wherever you go Under the popular. The number of people traveling abroad each year in our country is increasing sharply now. South......Tips & Frequently Asked Questions:Question 1: How long will the true-PDF of WS/T 191-2017_English be delivered?Answer: Upon your order, we will start to translate WS/T 191-2017_English as soon as possible, and keep you informed of the progress. The lead time is typically 1 ~ 3 working days. The lengthier the document the longer the lead time.Question 2: Can I share the purchased PDF of WS/T 191-2017_English with my colleagues?Answer: Yes. The purchased PDF of WS/T 191-2017_English will be deemed to be sold to your employer/organization who actually pays for it, including your colleagues and your employer's intranet.Question 3: Does the price include tax/VAT?Answer: Yes. 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