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WS 296-2017 English PDF

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WS 296-2017: Diagnosis for measles
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WS 296: Evolution and historical versions

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WS 296-2017English479 Add to Cart 4 days [Need to translate] Diagnosis for measles Valid WS 296-2017
WS/T 296-2017EnglishRFQ ASK 3 days [Need to translate] (Measles diagnosis) WS/T 296-2017
WS 296-2008English759 Add to Cart 4 days [Need to translate] Diagnostic criteria for measles Obsolete WS 296-2008

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Standard similar to WS 296-2017

GB 15979   GB/T 15981   WS/T 641   WS 295   WS 293   

Basic data

Standard ID WS 296-2017 (WS296-2017)
Description (Translated English) Diagnosis for measles
Sector / Industry Health Industry Standard
Classification of Chinese Standard C59
Word Count Estimation 19,124
Date of Issue 2017-07-24
Date of Implementation 2018-02-01
Older Standard (superseded by this standard) WS 296-2008
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 296-2017: Diagnosis for measles

---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.
(Measles diagnosis) ICS 11.020 C 59 WS People's Republic of China health industry standards Replacing WS 296-2008 Measles diagnosis Diagnosis for measles 2017 - 07 - 24 Posted 2018 - 02 - 01 implementation People's Republic of China National Health and Family Planning Commission released Directory Foreword ..II 1 Scope 1 2 Abbreviations .1 3 diagnostic basis .1 3.1 Epidemiological history .1 3.2 Clinical manifestations 1 3.3 laboratory testing .1 4 diagnostic principles .2 5 diagnosis 2 5.1 suspected cases 2 5.2 Clinical diagnosis of cases .2 5.3 Laboratory confirmed cases 2 5.4 exclude cases 2 6 differential diagnosis .2 Appendix A (normative) serological diagnostic methods .3 Appendix B (Normative) etiological diagnosis method .7 Appendix C (informative) Measles etiology, epidemiology and clinical manifestations .11 References .15

Foreword

Chapter 5 of this standard is mandatory, the rest are recommended terms. This standard was drafted in accordance with the rules given in GB/T 1.1-2009. This standard replaces WS 296-2008 "measles diagnostic criteria." This standard from the date of implementation, WS 296-2008 abolished at the same time. This standard compared with WS 296-2008, the main changes are as follows. - Modified abbreviations (see Chapter 2, Chapter 2 of the.2008 edition); - revised the diagnosis of measles (see Chapter 3, Chapter 3 of the.2008 edition); - revised the definition of clinical diagnosis (see Chapter 5, Chapter 5 of the.2008 edition); - Increased standard operation of laboratory test methods (see Appendix A and Appendix B,.2008 edition of Appendix A and Appendix B); - Increased measles epidemiology, especially within the epidemiological characteristics, clinical manifestations, complications, etc., of transmission of measles under the new situation (See Appendix C,.2008 edition of Appendix C). This standard was drafted. China Center for Disease Control and Prevention, China Center for Disease Control and Prevention virus prevention and control, the Capital Medical University Affiliated to Beijing Ditan Hospital, Capital Medical University Affiliated Beijing Children's Hospital, Guangdong Provincial Center for Disease Control and Prevention, Zhejiang Province Disease Prevention and Control System Center. The main drafters of this standard. Luo Huiming, Xu Wenbo, Yu Wenzhou, Li Xingwang, Ma Chao, Zhang Yan, Xie Zhengde, Hao Li Xin, Zheng Huizhen, Xie Shuyun, Su Qi Ru. This standard replaces the standards previously issued as follows. - GB 15983-1995; - WS 296-2008. Measles diagnosis

1 Scope

This standard specifies the diagnosis of measles, diagnostic principles, diagnosis and differential diagnosis. This standard applies to all types of medical and health institutions at all levels and their medical staff on the diagnosis of measles.

2 Abbreviations

The following abbreviations apply to this document. CPE. cytopathic effect DEPC. diethyl pyrocarbonate ELISA. enzyme-linked immunosorbent assay IgG. immunoglobulin G IgM. immunoglobulin M RF. rheumatoid factor RNA. ribonucleic acid RT-PCR. reverse transcription-polymerase chain reaction RPM. revolutions per minute OD. optical density value VTM. virus transportation medium

3 diagnostic basis

3.1 Epidemiology history 3.1.1 7 to 21 days before rash and measles patients diagnosed with contact history. 3.1.2 Before the rash 7 d ~ 21 d have measles epidemic areas of living or travel history. 3.2 Clinical manifestations 3.2.1 fever, body temperature is generally ≥ 38 ℃. 3.2.2 in the course of the first 3 days to 4 days began red rash, rash skin normal. Rash sequence generally from the ear, the face began, Expands from top to bottom and can affect mucous membranes. Rash time generally lasts 3 d ~ 5 d. 3.2.3 cough, runny nose, sneezing and other upper respiratory tract symptoms of catarrhal, and photophobia, tearing, conjunctivitis symptoms. 3.2.4 Early onset (usually in the course of the first 2 days to 3 days) in the oral buccal mucosa seen measles mucosal spots (Koplik spot). 3.3 laboratory testing 3.3.1 8 h to 56 d before blood sampling did not inoculate attenuated live attenuated measles vaccine, and blood samples within 28 d after measles IgM positive (See Appendix A). 3.3.2 Measles virus positive or isolated from measles virus in throat swabs or urine specimens (see Appendix B). 3.3.3 recovery of blood samples of measles IgG antibody titers than the acute phase ≥ 4-fold increase, or acute antibody negative and convalescent antibody positive transfer (See Appendix A).

4 diagnostic principles

According to epidemiological history, clinical manifestations and laboratory test results to be diagnosed.

5 diagnostic

5.1 suspected cases Available in 3.2.1, 3.2.2 and 3.2.3. 5.2 Clinical diagnosis of cases Suspected case of any of the following. a) have 3.1.1 and/or 3.1.2 and are not definitely diagnosed as other diseases; b) have 3.2.4; c) No specimens were collected for laboratory testing and no other diagnosis was clearly identified. 5.3 laboratory confirmed cases Suspected cases have any one of 3.3.1, 3.3.2, 3.3.3. 5.4 exclude cases Suspected case of any of the following. a) Measles IgM antibodies were negative for blood samples collected within 4 to 28 days after rash and did not comply with 3.1.1 and/or 3.1.2; b) Blood samples collected within 3 days after rash were tested for measles IgM antibody negative, measles virus nucleic acid negative in eligible throat swabs/urine samples Sexual, and does not comply with 3.1.1 and/or 3.1.2; c) No samples were collected for laboratory testing but were definitely diagnosed as other diseases; d) etiological specimens were isolated and identified as measles vaccine strain virus or vaccine virus positive for the virus, and no measles wild virus was also isolated Rash wild virus nucleic acid-positive; or in line with the following five cases. 1) a rash, with or without fever, but no cough and other respiratory symptoms; 2) rash after 7 to 14 days after inoculation of attenuated live vaccine containing measles; 3) Blood samples were collected from 8 d to 56 d after inoculation of live attenuated measles vaccine, and IgM was detected in measles. 4) epidemiological survey found no cases of recurrence caused by the case; 5) There are no other reasons that can be clearly explained by epidemiological and laboratory investigations.

6 differential diagnosis

The disease should be rubella, scarlet fever, children and other rash fever rash disease identification (see Appendix C). Appendix A. (Normative) Serological diagnostic methods A.1 Blood specimen collection, storage and transport A.1.1 Blood specimen collection Blood samples were taken within 28 days of rash for serological detection of measles IgM and IgG. A.1.2 In the following cases need to collect the second blood specimen Blood samples collected within 3 days after rash were negative for measles IgM antibody and no pharyngeal swabs/urine samples were collected for 4 d to 28 d The second blood specimen. A.1.3 blood sample collection process A.1.3.1 blood collection without anticoagulant 2 mL ~ 3 mL venous blood, marking on the tube wall. A.1.3.2 Blood samples can not be frozen, centrifuged at 1000 RPM for 10 min for serum separation; if centrifuge free, blood samples should be refrigerated 4 ℃ Place until serum is completely precipitated. A.1.3.3 Carefully aspirate serum, avoid aspiration of erythrocytes, under aseptic conditions, move to a serum tube with an external screw cap, do the tube wall Good mark. A.1.3.4 Fill out the specimen submission form, including the date of the last vaccination with the measles component, the date of the rash and the date of specimen collection. Sent Table to indicate the case number. A.1.4 storage of blood samples A.1.4.1 Blood samples should be stored refrigerated (2 ℃ ~ 8 ℃) and delivered to the testing laboratory within 24 hours. If you can not meet the above requirements, you should press A.1.3 method according to the separation of serum. A.1.4.2 sterile serum should be transported within 48 hours using ice boxes to the testing laboratory, or stored at 2 ℃ ~ 8 ℃, the maximum can not exceed 7 d. Long-term preservation of serum should be below -20 ℃ frozen, to avoid repeated freezing and thawing. A.1.5 transport of blood samples Specimens should be transported to the testing laboratory as soon as possible, will be filled with specimens of the container on a sealed plastic bag, with a cold box or thermos bottle delivery, The specimen inspection order on the plastic bag, and then tape fixed in the upper part of the freezer, transport date is confirmed, notify the receiving laboratory specimens Shipping time and shipping method. A.2 laboratory testing A.2.1 measles IgM antibody detection A.2.1.1 Purpose Is mainly used for qualitative and/or quantitative detection of anti-measles virus in human serum or plasma IgM antibodies for early diagnosis of measles. A.2.1.2 operation steps (or according to the reagent manual) A.2.1.2.1 Quantitative ELISA method for the detection of measles virus IgM antibody sample dilution method A.2.1.2.1.1 10 μL of sample to be tested should be diluted with 500 μL of dilution buffer to prepare a patient sample dilution. A.2.1.2.1.2 Add 40 μL of RF to 100 μL of patient sample diluent (A.2.2.2.1.1) and use 60 μL of dilution buffer Line dilution, the final specimen dilution ratio of 1.100. A.2.1.2.2 Test procedure A.2.1.2.2.1 Place the required number of microporous strips on the microplate frame and prepare a label. A.2.1.2.2.2 Add RF test serum samples, adsorption at room temperature 15 min or 4 ℃ overnight. A.2.1.2.2.3 Add 100 μL of diluted sample or ready-to-use control serum to each well. Leave a hole as a substrate empty White, for example. Quantitative IgM antigen micropore Hole A1 Hole B1 Hole C1 Hole D1 Hole E1 Substrate blank Negative control Standard serum Standard serum Specimen 1 A.2.1.2.2.4 Incubate the sample in a wet box (37 ± 1) ° C for (60 ± 5) min. A.2.1.2.2.5 After incubation, wash wells with wash buffer (use automatic plate washer or hand wash). Sip or rinse off the lotion first; then Add 300 μL wash solution to each well, and then suck or rinse the wash solution, repeating the washing process for a total of 4 times. Finally, flip the microplate over the paper towel Dry, so that no longer contain liquid pore. A.2.1.2.2.6 adding enzyme-labeled antibody. In the appropriate hole (except the substrate blank wells) added 100 μL IgM enzyme-labeled antibody, wet box (37 ± 1) ℃ for 30 ± 1 min. A.2.1.2.2.7 Wash wells with wash buffer (see A.2.2.2.2.5). A.2.1.2.2.8 Add Substrate. Add 100 μL of substrate solution (including blank wells) to each well and incubate (37 ± 1) ° C in a humid chamber (30 ± 1) min. A.2.1.2.2.9 Stop the reaction. Add 100 μL stop solution to each well and gently agitate the microplate to mix the solution. A.2.1.2.2.10 read the OD value. Substrate blank hole blank control, OD value was read within 60 min. A.2.1.3 quality control standards A.2.1.3.1 Blank OD value of the substrate must be < 0.25. A.2.1.3.2 Negative controls must be negative. A.2.1.3.3 The average OD of the standard serum must be within the valid range given in the kit-specific quality control certificate (minus Go to the substrate blank). A.2.1.3.4 If the above criteria are not met, the test is invalid and should be repeated. A.2.1.4 Evaluation of the results According to the standard serum OD value calculated CUT-OFF value of the upper and lower limits, the specimen OD value ≥CUT-OFF value of the upper limit, the result was judged as positive; specimen OD value < CUT-OFF value of the lower limit, the result was judged as negative; in the sample OD value between CUT-OFF value of the upper and lower limits, the results were judged as suspicious. According to the standard curve provided by the kit, calculate the antibody activity value in the specimen. A.2.2 ELISA assay measles IgG antibody A.2.2.1 Use Mainly used for qualitative and/or quantitative detection of human serum or plasma anti-measles virus IgG antibodies for measles diagnosis or human antibody levels survey. A.2.2.2 operation steps (or according to the reagent manual) A.2.2.2.1 Quantitative ELISA method for the detection of measles virus IgG antibody sample dilution method 1000 μL of dilution buffer should be used to dilute the sample to be tested in 10 μL to prepare a patient sample dilution and a final sample dilution Release ratio of 1.100. A.2.2.2.2 Test procedure A.2.2.2.2.1 Place the required number of microporous strips on the microplate frame and prepare a label. A.2.2.2.2.2 Add 100 μL of diluted sample or ready-to-use control serum to each well. Leave a hole as a substrate empty White, for example. Quantitative IgM antigen micropore Hole A1 Hole B1 Hole C1 Hole D1 Hole E1 Substrate blank Negative control Standard serum Standard serum Specimen 1 A.2.2.2.2.3 Incubate the sample in a wet box (37 ± 1) ° C for (60 ± 5) min. A.2.2.2.2.4 After incubation, wash wells with wash buffer (use automatic plate washer or hand wash). Sip or rinse off the lotion first; then Add 300 μL wash solution to each well, and then suck or rinse the wash solution, repeating the washing process for a total of 4 times. Finally, flip the microplate over the paper towel Dry, so that no longer contain liquid pore. A.2.2.2.2.5 Enzyme-labeled antibody. Add 100 μL of IgG enzyme-labeled antibody into the corresponding wells (except for blank wells) (37 ± 1) ℃ for 30 ± 1 min. A.2.2.2.2.6 Wash plate wells with wash buffer (see A.2.2.2.2.5). A.2.2.2.2.7 Add Substrate. Add 100 μL of substrate solution (including substrate blank wells) to each well and incubate (37 ± 1) ° C in a humid chamber (30 ± 1) min. A.2.2.2.2.8 Stop the reaction. Add 100 μL stop solution to each well and gently agitate the microplate to mix the solution. A.2.2.2.2.9 read the OD value. blank control substrate blank control, OD value was read within 60 min. A.2.2.3 Quality control standards A.2.2.3.1 Blank OD value of the substrate must be < 0.25. A.2.2.3.2 Negative controls must be negative. A.2.2.3.3 The average OD of the standard serum must be within the valid range given in the kit-specific quality control certificate (minus Go to the substrate blank). A.2.2.3.4 If the above criteria are not met, the test is invalid and must be repeated. A.2.2.4 Evaluation of the results According to the standard serum OD value calculated CUT-OFF value of the upper and lower limits, the specimen OD value ≥CUT-OFF value of the upper limit, the result was judged as positive; specimen OD value < CUT-OFF value of the lower limit, the result was judged as negative; in the sample OD value between CUT-OFF value of the upper and lower limits, the results were judged as suspicious. According to the standard curve provided by the kit, calculate the antibody activity value in the specimen. AA

Appendix B

(Normative) Etiological diagnosis B.1 virus isolation B.1.1 specimen collection and processing B.1.1.1 specimen collection Specimens for virus isolation include throat swabs and urine. The measles cases should be rash before 5 days to 5 days after rash to take the above specimens. B.1.1.2 specimen handling methods B.1.1.2.1 throat swab specimens Sterile cotton swab slightly dipped in saline, repeated smear the patient's throat several times, and then the cotton swab immersed in 1 mL ~ 2 mL of sample maintenance solution (containing 500 to 1000 units/mL penicillin, 500 μg/mL to 1000 μg/mL streptomycin and 2% bovine serum DMEM) or VTM, and repeatedly squeezed, cotton swab abandoned. Specimen maintenance solution is best to add non-cytotoxic nystatin 50 μg/mL or amphotericin 5 μ g/mL, placed in 4 ℃ overnight before inoculation of cells for inactivation of fungi. B.1.1.2.2 Urine specimens Aseptically collect 30 mL ~ 50 mL middle urine in 50 mL screw-cap sterile plastic centrifuge tube, refrigerated to the laboratory as soon as possible, 2 000 Rpm, centrifuged for 5 min, the supernatant was discarded, the pellet was resuspended with 2 mL of sample maintenance solution or VTM, and the resuspended sample solution was frozen at -70 ° C; Non-centrifuged urine should not be frozen. B.1.2 specimen inoculation The above treated samples need at least -70 ℃ freeze-thaw once before inoculation to facilitate the release of virus from epithelial cells. Take 0.3 mL ~ 0.5 mL of the sample solution is inoculated into a well-grown monolayer of passage cell lines such as African green monkey kidney cell/lymphocyte signaling activator-transfected Africa Vero cell transfected to express the human signaling lymphocyte activation molecule Vero/SLAM), and the solution was stored at 37 ° C for 1 h to 2 h before discarded. The solution was added to maintain the solution at 37 ° C. The next day, cytotoxicity was observed. Change fluid if necessary. B.1.3 observe the CPE Observed CPE daily progress after inoculation, continuous observation of 7d, if there is a characteristic measles virus CPE appeared observed until 75% Cytopathogenic lesions (CPE), frozen at -70 ° C. If there is no CPE, which set -70 ℃ frozen 3 times, and then blind pass 1 to 2 generations. B.1.4 Identification of strains Measles virus strains can be identified using fluorescent quantitative RT-PCR, RT-PCR and sequencing (see Appendix B.2). B.2 Virus Nucleic Acid Test B.2.1 specimen collection and processing Refer to B.1.1 for specimen collection and processing. In the delivery of specimens, pay attention to low-temperature transport conditions, sent to the specimen in the experiment After the room, do not repeatedly frozen and thawed specimens, so as to prevent nucleic acid degradation. B.2.2 Virus nucleic acid extraction Using commercial kits or automated nucleic acid extractors, refer to kit instructions and/or automated nucleic acid extraction instrument instructions With instructions. You can also use TRIZOL extraction, the specific steps are as follows. a) Add 250 μL virus suspension to a 1.5 mL centrifuge tube, then add 750 μL TRIZOL solution, mix for 5 s and centrifuge. b) Add.200 μL of chloroform solution, mix well for 5 s; c) Allow to stand at room temperature for 10 min and then centrifuge at 10,000 RPM for 20 min at room temperature. d) Transfer the supernatant to a new 1.5 mL centrifuge tube; e) then add 500 μL isopropanol solution, shake 10 s; f) Let stand at room temperature for at least 15 min; g) Centrifuge at 13 000 RPM for 25 min at 4 ° C; h) Discard the supernatant and add 950 μL of ice-cold 70% ethanol; i) Centrifuge at 13,000 RPM for 20 min at 4 ° C; j) Carefully aspirate the supernatant with a tip; k) After drying at room temperature, add 15 μL of DEPC-treated H 2 O and add 20 U of RNasin (RNase protein inhibitor) -20 ℃ save. Note. The entire operation with masks and disposable gloves. B.2.3 Fluorescence quantitative RT-PCR method to detect measles virus nucleic acid B.2.3.1 reaction system Ingredient volume/μL DEPC water 3.5 Reaction Buffer 12.5 Upstream primer (7.5 μM) 1.0 Downstream primer (7.5 μM) 1.0 Probe (2.5 μM) 1.0 DNA polymerase 0.5 Reverse transcriptase 0.5 Template nucleic acid 5.0 Total 25.0 B.2.3.2 Primer and probe sequences B.2.3.2.1 Upstream primer sequence. 5'-TGG CAT CTG AAC TCG GTA TCA C-3 ' B.2.3.2.2 Downstream primer sequences. 5'-TGT CCT CAG TAG TAT GCA TTG CAA-3 ' B.2.3.2.3 Probe Sequence. 5 '(FAM) -CCG AGG ATG CAA GGC TTG TTT CAG A-3' (BHQ) B.2.3.3 reaction conditions 42 ℃ ••••••••••••••••••• 5 min 95 ℃ ••••••••••••••••••• 10 s 95 ℃ •••••••••••••••••••••••••• 5 s × 40 cycles 55 ℃ ••••••••••••••••••• 30 s Note. Fluorescent labeling and collection time refer to kit instructions. B.2.3.4 Results Analysis and Judgment Results analysis and judgment method reference kit instructions. B.2.4 measles virus identification and genotyping target gene amplification and sequencing B.2.4.1 Amplification of N-terminal carboxy-terminal 634 nucleotide sequences B.2.4.1.1 Reaction system Ingredient volume/μL DEPC water 7.5 Reaction Buffer 12.5 Upstream primer (20 μM) 0.5 Downstream primer (20 μM) 0.5 Reverse transcription and DNA polymerase 1.0 Template nucleic acid 3.0 Total 25.0 B.2.4.1.2 Primer sequences B.2.4.1.2.1 Upstream primer sequence. 5'-TGG AGC TAT GCC ATG GGA GT-3 ' B.2.4.1.2.2 Downstream primer sequences. 5'-TAA CAA TGA TGG AGG GTA GG-3 ' B.2.4.1.3 The reaction was carried out in a PCR machine as follows 50 ℃ ••••••••••••••••••• 30 min 94 ℃ ••••••••••••••••••• 2 min 94 ℃ •••••••••••••••••••• 30 s 55 ℃ ••••••••••••••••••••••••• 30 s × 30 cycles 72 ℃ ••••••••••••••••••• 1 min 72 ℃ ••••••••••••••••••• 10 min 4 ℃ ••••••••••••••••••••• Save B.2.4.1.4 Detection and identification of PCR products B.2.4.1.4.1 Preparation 1% ~ 2% agarose gel Take 100 mL electrophoresis buffer (1 × TAE) into a clean flask, add 1 g ~ 2 g agarose powder, gently shake the triangle Bottle, the agarose particles are uniformly cloudy state; microwave heating agarose melting; melted agarose naturally cooled to about 70 ℃, Pour into prepared gel bed, the gel thickness is about 0.3 cm ~ 0.5 cm; standing at room temperature, gel curing. B.2.4.1.4.2 Gel electrophoresis The prepared agarose gel was placed in the electrophoresis tank; electrophoresis buffer was added electrophoresis buffer (1 × TAE), the amount of buffer to Exceed the surface of the gel 1 mm ~ 2 mm is appropriate; sample pipette 5 μL with the sampler, mix with the sample buffer, gently added to the gel sample Hole in the product; voltage selection 100 V, start electrophoresis. When the indicator has migrated to the proper place, turn off the power and remove the gel; place the gel in Containing ethidium bromide (EB) solution, staining for about 10 min; gel electrophoresis camera electrophoresis strip, save the record. Other products can also be used The dye instead of EB, the use of reference dye instructions. B.2.4.1.5 gel cut PCR products purified The PCR product was purified using a kit. Specific steps see kit instructions. B.2.4.1.6 Mark reaction B.2.4.1.6.1 Preparation of labeled reaction system Big Dye 2 μL Reaction buffer 2 μL Primer (4 μM) 1 μL Template (purified PCR product) 2 μL ~ 8 μLa Water 20 μL a According to the purpose of the degree of bright strip to choose the amount. B.2.4.1.6.2 Marker reaction in the PCR machine according to the following procedure 96 ℃ •••••••••••••••••••• 1 min 96 ℃ ••••••••••••••••••• 10 s 50 ℃ •••••••••••••••••••••••••• 5 s × 25 cycles 60 ℃ ••••••••••••••••••• 4 min 4 ℃ ••••••••••••••••••••• Save B.2.4.1.6.3 labeled reaction product was purified Weigh 2.7 g of G-50 powder into a 50 mL centrifuge tube and add deionized water to 50 mL. After mixing thoroughly, let stand at room temperature for 30 min, So that full hydration; to the spin column by adding mixed G-50 suspension 900 μL, allowed to stand for 10 min; pressure, the excess moisture from Out of the bottom of the stem; then place the spin column on a 2 mL collection tube, centrifuge at 3,000 RPM for 2 min at room temperature; transfer the spin column to In a 1.5 mL centrifuge tube, add the labeled product to the center of the bevel of the gel. Centrifuge at 3,000 RPM for 2 min at room temperature and collect the purified label product. B.2.4.1.7 Sequence determination The purified labeled product was added to a 96-well PCR plate, enter the sample number in the computer, and then follow the sequence analyzer Instructions for follow-up operation. B.2.4.1.8 Sequence collation and analysis The sequence data obtained from the sequencer is saved as a standard graphic file (.ab1) and the sequence is collated using the relevant sequence analysis software Pieces of the sequence were edited, as well as genetic relationship, nucleotide and amino acid homology analysis. BB

Appendix C

(Informative) Etiology, epidemiology and clinical manifestations of measles C.1 Measles etiology Measles virus is the causative agent of measles and was first obtained from measles patients in the 1950s by John Enders and Thomas Peebles Liquid separation. The measles virus is one of the most contagious pathogens known and humans are the only natural hosts of the measles virus. The measles virus is an enveloped, single-stranded RNA virus belonging to the genus Paramyxoviruses. The whole genome of measles virus, N. (Nucleoprotein, nuclear protein) and H (Hemagglutlutin, hemagglutinin) gene mutation is relatively large, which N-carboxy-terminal 450 nucleotide variation of the largest, this sequence can identify both measles virus, and measles virus gene-targeted target sequence. Through the N-carboxy Base...

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