, (WP)IICPIIVD/I.I/OO I-A RS/2006/GE/OS(HOK) English only
REPORT HANDS-ON TRAINING ON THE LABORATORY DIAGNOSIS OF MEASLES Kowloon, Hong Kong (China) 13-18 March 2006
Convened by: WORLD HEALTH ORGANIZATION REGIONAL OFFICE FOR THE WESTERN PACIFIC
Not for sale Printed and distributed by: World Health Organization Regional Office for the Western Pacific Manila, Philippines
WHOIWPRO LIBRAR\, MANILA. PHILIPPINES
2 2 1~11'~' ?D05
NOTE The views expressed in this report are those of the participants in the Hands-on Training on the Laboratory Diagnosis of Measles and do not necessarily reflect the policies of the World Health Organization. The Expanded Programme on Immunization, WHO Western Pacific Regional Office, would like to thank the Virology Division, Public Health Laboratory Services Branch, Centre for Health Protection, Department of Health, Hong Kong (China), for hosting and providing resource persons and technical support for the workshop, and the Centers for Disease Control and Prevention (CDC), Atlanta, and the National Institute for Immunizations and Infectious Disease, Tokyo, for providing financial and technical support for the meeting including the p·roduction of this document.
Keywords: . Measles laboratory testing, IgM ELISA, DBS (dried blood spots), quality assurance (QA)
This report has been printed by the Regional Office for the Western Pacific of the World Health Organization for the participants in the Hands-On Training on The Laboratory Diagnosis of Measles, 13-18 March 2006
SUMMARY
A training on laboratory diagnosis of measles virus infection was held at the Virology Division, Public Health Laboratory Services Branch, Centre for Health Protection, Department of Health, Hong Kong (China) from 13 to 18 March 2006. Participants from six national measles laboratories attended the training. In addition to support from the WHO secretariat, the workshop was conducted with support from temporary advisers from the Virology Division, Public Health Laboratory Services Branch, Centre for Health Protection, Department of Health, Hong Kong (China); Center for Disease Control and Prevention (CDC), Atlanta; and National Institute for Immunizations and Infectious Disease, Tokyo. The objectives of the workshop were: (1)
to introduce WHO standardized methods for laboratory diagnosis of measles infection;
(2) to familiarize participants with practical procedures of measles immunoglobulin class M (IgM) enzyme immunoassays (EIAs) of serologic confirmation of acute measles infection; (3) to familiarize the participants with the techniques for isolation of measles virus using the Vero/SLAM cell line; (4) to introduce the participants to the use of molecular epidemiologic techniques to study the transmission of measles virus; (5) to familiarize participants with laboratory quality assurance inclusive of standard operating procedures and internal quality control; (6) to introduce the participants to the WHO network laboratory accreditation scheme and its criteria to be met; and (7) to introduce laboratory data management and reporting to the Western Pacific Regional Office. The training consisted of presentations by course facilitators, brief presentations by participants of each country, laboratory practices and demonstrations. Overall, the training was extremely successful. The staff of the Virology Division did an outstanding job in preparation for the course. The participants showed a high level of technical competence. The participants were encouraged to remain in contact with each other and with the facilitators as a future resource for technical advice and support.
CONTENTS
Page
1. INTRODUCTION ....................................................................................................... I 1.1 Objectives .......................................................................................................... I 1.2 Organization .... .................................................................................................. I
2. PROCEEDINGS .......................................................................................................... 2 2. I Presentations ...................................................................................................... 2 2.2 Practical sessions ............................................................................................... 2
3. CONCLUSIONS ........................................................................................................ 3
ANNEXES: ANNEX 1 ANNEX 2
LIST OF ATTENDEES PROGRAMME SCHEDULE
I. INTRODUCTION
The Western Pacific Region ofthe World Health Organization (WHO) has declared a goal of regional measles elimination with a target date of2012, recommended by a Measles Task Force in July 2004 and a Technical Advisory Group meeting in June 2005. The Regional Committee for the Western Pacific considered this target date at its fifty-sixth session in September 2005. WHO recommends that the clinical diagnosis of measles be confirmed by the detection of measles immunoglobulin class M (IgM) using an enzyme immunoassay (EIA). To increase familiarity of some laboratory staff in the Region to measles IgM EIA testing and the use of dried blood samples as an alternative method of sampling, the WHO Western Pacific Regional Office held a training course at the Virology Division, Public Health Laboratory Services Branch, Centre for Health Protection, Department of Health, Hong Kong (China). 1.1 Objectives (I)
To introduce WHO standardized methods for laboratory diagnosis of measles infection.
(2) To familiarize participants with practical procedures of measles immunoglobulin-class M (IgM) enzyme immunoassays (EIAs) of serologic confirmation of acute measles infection. (3) To familiarize the participants with the techniques for isolation of measles virus using the Vero/SLAM cell line. To introduce the participants to the use of molecular epidemiologic (4) techniques to study the transmission of measles virus. (5) To familiarize participants with laboratory quality assurance inclusive of standard operating procedures and internal quality control. (6) To introduce the participants to the WHO network laboratory accreditation scheme and its criteria to be met. (7) To introduce laboratory data management and reporting to the Western Pacific Regional Office. 1.2 Organization
The 10 participants represented six countries in the Western Pacific Region: China, Malaysia, Mongolia, the Philippines, the Republic of Korea, and Viet Nam. In addition to support from the WHO secretariat, the training was conducted with support of three temporary advisers from the Virology Division, Public Health Laboratory Services Branch, Centre for Health Protection (CHP), Department of Health, Hong Kong (China); Center for Disease Control and Prevention (CDC), Atlanta; and National Institute for Immunizations and Infectious Disease (NIID), Tokyo (Annex I).
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2. PROCEEDINGS
2.1
Presentations
The participants heard 13 presentations from the course facilitators (see timetable, Annex 2). The presentations described the role of the laboratory in surveillance for measles and rubella. Emphasis was placed on the importance of serologic testing, virus isolation techniques using the Vero/SLAM cell line, the importance of genetic characterization of circulating strains of measles and rubella viruses, the critical importance of quality control in the laboratory, data analysis, and data management. In addition to the presentations made by the course facilitators, one representative from each country gave a brief presentation of the status of measles control in his/her country and each gave a description of the organization and activities ofthe national measles laboratory. 2.2 Practical sessions
The training included five laboratory practice exercises consisting of performing the DadeBehring EIA for measles IgM, the Dade-Behring EIA for rubella IgM, the Denka Seikan EIA for measles IgM, virus isolation using Vero/SLAM cells, and immunofluorescence to detect measles virus in cell culture. The participants also observed demonstrations of passaging Vero/SLAM cells and calibration of pipettes. All participants perfonned three EIA assays during the training, observed measles cytopathic effect (CPE) following inoculation ofVero/SLAM cells, and performed immunoflfuorescence testing. Following the completion of each exercise, there was an open discussion between the facilitators and the participants. During the laboratory exercises, there were always three to five facilitators present to assist the 10 participants. The facilitators were: Dr Kazunobu Kojima, WHO, Regional Office for the Western Pacific, Manila Dr Wilina Lim, CHP, Hong Kong (China) Dr Kei Numazaki, NIID, Tokyo Dr Paul Rota, CDC, Atlanta Dr Andrew Leung, CHP Mr KC Szeto, CHP Ms Wong Wai Man, CHP Ms Wong Kit Lee, CHP Ms Lee Wai Yan, CHP Mr CS Lau, CHP Ms Mary Lai, CHP Ms SChow, CHP The facilitators prepared a panel of 44 serum samples for each group of participants, organized laboratory equipment for each group, and prepared a package containing copies of all of the laboratory protocols and data sheets. This outstanding preparation allowed the students to complete the laboratory exercises in a very efficient manner without unnecessary delays or confusion. In addition, the facilitators from the Virology Section gave detailed instructions to the participants, observed their laboratory practices, and assisted with calculation and interpretation of the results.
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The results from the EIA assays were remarkably consistent and only a few inconsistent results were noted. Each participant tested 44 samples, unprecedented for a WHO laboratory training course. Each participant completed an examination consisting of25 review questions. The results of the examination were then discussed. All participants received passing scores.
3. CONCLUSIONS
Overall, the course was extremely successful. The facilities within the Virology Section are outstanding. Three laboratories were dedicated to the training, providing ample space for all ofthe participants. The conference room used for the presentations was state-of-the art. Participants had access to three biological safety cabinets, and all of the necessary equipment was in excellent condition and available in the training laboratories. The staff of the Virology Section did an outstanding job in preparation for the course. The participants showed a high level of technical competence. All participants completed course evaluation sheets, which are on file. Participants were given Vero/SLAM cells, Geneticin, and Denka-Seikan EIA kits to take back to their laboratories.
-4ANNEXl
WORLD HEALTH ORGANIZATION
ORGANISATION MONDIALE DE LA SANTE
REGIONAL OFFICE FOR THE WESTERN PACIFIC BUREAU REGIONAL DU PACIFIQUE OCCIDENTAL
HANDS-ON TRAINING ON THE LADORA TORY DIAGNOSIS OF MEASLES Kowloon, Hong Kong 13-18 March 2006
2 March 2006 ENGLISH ONLY
LIST OF PARTICIPANTS, TEMPORARY ADVISERS AND SECRETARIAT 1. PARTICIPANTS
CHINA
Dr MAO Naiying, Medical Officer - Assistant Researcher, Chinese Center for Disease Control and Prevention, 27 Nanwei Lu Road, Beijing 100050, Tel. No. (8610) 6302 7904, Fax No. (8610) 6302 8480, E-mail: mecho429@sohu.com Mr MOHD APANDI bin Yusof, Research Officer, Virology Unit, Institute of Medical Research, Ialan Pahang, 50588 Kuala Lumpur, Tel No. (603) 2698 6033/013 845 3167, Fax No. (603) 2693 8094, E-mail: apandi@imr.gov.my Dr/Ms MYAGMARSUREN Shijirbaatar, Virologist, National Reference Laboratory, National Centre for Communicable Diseases, Ulaanbaatar-13, Post Box 0583, Tel. No. 976 11455815 or 99735350, E-mail: miga_sh@yahoo.com Ms Bibiana P. PASCUA, Medical Laboratory Technician I, Virology Department Research Institute for Tropical Medicine, Department of Health, Filinvest Corporate City Compound, Alabang, Muntinlupa City, Telefax. No. (632) 809 7120"E-mail: Ms Ligaya C. PICAZO, Science Research Specialist II, Virology Department Research Institute for Tropical Medicine, Filinvest Corporate City Compound, Alabang, Muntinlupa City, Tel. No. (632) 809 7120, Fax No. (632) 809 7120, E-mail: 1cpicazo@yahoo.com
MALAYSIA
MONGOLIA
PHILIPPINES
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"
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REPUBLIC OF KOREA
Dr CHUNG Yoon-Seok, Researcher, Center for Infectious Disease Research, National Institute of Health, Korea Centre for Disease Control Prevention, 5, Nokbun-dong Eunpyung-gu, Seoul 122-701, Tel. No. 8223801504, Fax No. 8223892014, E-mail: cys93@nih.go.kr DrlMs NGUYEN Thi Hien Thanh, National Institute of Hygiene and Epidemiology, No.1 Yersin Street, Ha Noi 10 000, Tel. No. 04 8211534, Fax No. 04 8 213 782, E-mail: thanhpoliolab@yahoo.com Dr/Ms PHAM Thi Thu Hang, National Institute of Hygiene and Epidemiology, No.1 Yersin Street, Ha Noi 10000, Tel No. 04 9715678, Fax No. 04 8213 782 E-mail:
VIETNAM
VIETNAM
Dr NGUYEN Thanh Long, Researcher, Pasteur Institute, 167, Pasteur Street, District 3, Ho Chi Minh City. Tel. No. (848) 8 202 878, Fax No. (848) 8 231 419, E-mail: ntlongdr@yahoo.com Ms PHAM Thi Nhung, Researcher, Pasteur Institute, Ho Chi Minh City, Tel. No. (848) 8 202 878, Fax No. (848) 8 231419, E-mail: saotim20@yahoo.com
2. TEMPORARY ADVISERS
Dr Paul ROTA, Microbiologist, Measles Virus Unit, Center for Disease Control and Prevention, Mailstop C-22, 1600 Clifton Road, Atlanta GA 30033, United States of America, Tel. No.1 4046394181, Fax No. 14046394187, E-mail: prota@cdc.gov Dr Wilina LIM, Consultant-Medical Microbiologist, Centre for Health Protection, Department of Health, 9/F Public Health Laboratory Centre, 382 Nam Cheong Street, Shek Kip Mei, Kowloon. Hong Kong (China), Tel. No. (852) 2319 8252, Fax No. (852) 2319 5989, E-mail: wllim@pacfiic.net.hk Dr Kei NUMAZAKI, Chief of Measles Laboratory, Virology III, National Institute ofInfectious Diseases, 4-7-1 Gakuen Musashi-murayama, Tokyo 208-00 II, Japan, Tel. No. +81 42 561 0071 (Ext. 707), Fax No. +81 42 565 3315, E-mail: numazaki@nih.go.jp
3. SECRETARIAT
Dr Kazunobu KOJIMA, Scientist (Laboratory Virologist), Expanded Programme on Immunization, World Health Organization, Regional Office for the Western Pacific, United Nations Avenue, 1000 Manila, Philippines, Tel. No. +632-528-9750, Fax No. +632-521-1036, E-mail:kojimak@wpro.who.int
HANDS-ON TRAINING ON THE LIIBORI\TORY DIAGNOSIS OF M(I\Sl.FS Hong Kong (China) 13-18 March 2006 TENTATIVE TIMETIIBLE Time Monday, 13 March REGISTRATION Time ----~
English only 18 February 2006
Tuesday, 14 March 5(0) Inti oduclioll 10 Dade Seilring 19M assay
Time
Wednesday, 15 March 5(a) • Laboratory practice: Dade Behring Rubella 19M assay (continuation)
08:30-08:45 08:45-09:30
08:30-08:45 08:45-10:00
08:30-10:00
1
09:30-10:00
2
· · · ·
Opening ceremony Opening remarks Self-introduction Administrative announcements
.
L"h(lr;olloIY practice: Dade S81111ng Measles IgM assay
Group photograph
10:00-10:30 10:30-11:00
Overview or progrmnmes (a) WHO Western Pacific Regional plmls for measles elimination and labnratory network ---Coffee break
-10:00-10:30 ~--
---Coffee break
10:00-10:30 10:30-12:00 5{a) •
Coffee break Laboratory practice: Dade Behring Rubella IgM assay (continuation) Calculation and interpretation of results 0\
(b) Country presentations on measles elimination and laboratory activities (10 mins) China MalAysia Mongolia Disclissions
10:30-12:30
11:00-11:15
·
· ·
5(a) • Lahoratory practice: Dade Behring Measles 19M assay (contimlRlion)
3. 11:15-1130 11:30-12:15 12:15-12:30 12:30-13:30 13:30-14:00 14:00-14:30 14:30-15:00 15:00-1515 15:15-15:45 15:45-16:00 16:00-16:15 16:15-16:30 16:30-16:45
Quality assurance (a) Good labomlory rraelices
I
4. Measles molecular epidemiology and role of virus isolation Discussions Lunch break
-- - - - - -
12:30-13:30 13:30~1600
Lunch break 5(8) laboratory pmclice: Dade Rehling Measles IgM assay (r.onfiflllali()n) Calculation and interpretation of results
12:00-13:00 15:00-16:00
Lunch break 2(a) Country presentations on measles elimination and laboratory activities (10 mins)
5. Introduction of IgM testing 3{a) • Specimen collection, handling and processing 3(b) Safely in the laboratory Discussion -
•
· · 15:30:16:00 16:00-17:00 17:00-17:30 cultures
•
Philippines Republic of Korea VietNam
~
Coffee break ~---
16:00~16:30
-
----------
Coffee break (He) I 8bomtqry practice" inoculAtion of cell r,I,ilurf?s
Coffee break Laboratory practice: observation of cell 5(c) Introduction to alternative sampling techniques
3(a) • (a) (b) Dinner
Inlroduction 10 the laboratory
16:30~1730
6. Introduction to coli culhtre pror.edures
VerolSLIlM cell line LAhoratory practice: cell cultllres, --------~~---~
y~roISLN~~r.,,1I plor_a9~tion ------
Z t"'.1 X N
~
~-'''''''''';.''-.~7"~7'''''''
~--.
Time
Thursda y. 16 March --~t
Time ----~~
..
08 :30-1 0 :00
5(b) Inlroduction to Denka Selken IgM a ss~y, and Laboratory pmr:tice: Denka Sp.iken IgM assay mE'asl~s
U8:~0-09:00
Frin"y. 17 March ---- ---- - ._- _._-_._. :1(1') • Introdllction of good d~la m~nagempnt Ann documentA.tion ~(c)
Time -
Saturda y, 18 March
> N
09:00-09 :30 09:30-10 :30
7.
Course evaluation
= = ~
Round table/Discussion (trouble shooting and individual consultations)
09:00-09 :30
• Intrndurtiron tn Wer;tnrn Pacific Region me~stes I"horatory oatClbase system and monthly reporting
. - -..- -
09:30·10 :00
Hnrvest crl1s :lod Ir Ip<;t ~_~ .(a~drP.p.e"t tf3."t.'n9 i~ "'.'1~i!!'?L ___ .
10:00·10 :30 11:00-12 :30
Coffee break 5(b) Laboratory practice: Denka Seiken measles IgM assay (continuation)
10:00·10 :30 10:30·12 :30
Coff"" break Harvest cells and IF test (continuation)
10:30-11 :00 11 :00-12:0 0 12:00 B.
Coffee break Conclusions Closing ceremony !
9.
-..J
12:30-13:30 13:30-15:00 15:00-16:00
Lunch break 5(b) Laboratory practice: Denkil Seiken measles IgM assay (contilluatioll) Calculation, review and interpretation of results of Denka Seiken tests
12:30-13 :30 13:30·14 :00 14:00·14 :30 14:30-16 :00 3
Lunch break Quality assuran ce (d) Shipping of "~mptes in th .. network Ex;:tm mview
I
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16:00-16:30 16:30-17:00 17:00-1730
Coffee break 51b) Laboratory practice: observation of cell cultures l.aboratory Quiz (overnight) -
16:00-16 :30 -
Coffee brE'IIk Exam review (colllim/illian)
16:30-17 :30