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Hong Kong’s “Children of 1997” Biobank Project 「九七的兒女」出生隊列生物庫

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Presentation on theme: "Hong Kong’s “Children of 1997” Biobank Project 「九七的兒女」出生隊列生物庫"— Presentation transcript:

1 Hong Kong’s “Children of 1997” Biobank Project 「九七的兒女」出生隊列生物庫
- Sample collection, processing and storage Cindy Lin 林詩琳 School of Public Health, The University of Hong Kong 3-Nov-14

2 “Children of 1997” Birth Cohort 「九七的兒女」出生隊列
8,327 infants born in April and May 1997, Hong Kong 7,154 active follow-up in Hong Kong currently 3-Nov-14

3 “Children of 1997” Birth Cohort 「九七的兒女」出生隊列
8,327 infants born in April and May 1997, Hong Kong 7,154 active follow-up in Hong Kong currently 3-Nov-14

4 “Children of 1997” Birth Cohort 「九七的兒女」出生隊列
8,327 infants born in April and May 1997, Hong Kong 7,154 active follow-up in Hong Kong currently 生物庫 3-Nov-14

5 Biobank Project What ? A bank of “biomaterials” (samples of blood, urine, stool and etc.) Why ? Chronic disease pattern Biomarkers Genetics and epigenetics Microbiome How ? Preparation work Sample collection Sample processing and storage 3-Nov-14

6 4. Schedule & Participant Recruitment
Preparation work 前期準備 基礎設施 標準作業程序 時間表 招募參與者 問卷設計 1. Infrastructure Facilities for sample processing Storage center Setting up clinics Inventory system for tracking samples Stock management system 2.Protocol Standard operating procedure (SOP) Sample transfer protocol 3. Questionnaire design ALSPAC cohort Validated instrument in local population 4. Schedule & Participant Recruitment Targeted area and number Invitation letter Follow-up calls 3-Nov-14

7 Pilot study 試驗性研究 There are always some problems you can not anticipate … Response Improve the participant response rate Utility Maximize the utility of available resources Time Streamline the overall procedures 3-Nov-14

8 currently 3 operating in parallel
(Aug 2013 – Oct 2014) NP YMT HKU PWH We have 4 clinics, currently 3 operating in parallel Blood sample processing site Other biomaterials processing site (Lab in School of Public Health, SPH) Storage center (Center for genomic sciences, CGS) 3-Nov-14

9 Stock inventory 物資庫存管理
3-Nov-14

10 SOP 標準作業程序 3-Nov-14

11 Principles for SOP 標準作業程序制定原則
Avoid processing approach that inherently preclude future analysis Serve as wide a range of scientific investigation as can be anticipated Future proofing Consistency Centralize and standardize processing methodology Quality control and assurance Ultra low temperature to ensure long term stability and integrity Protect samples from freeze thaw degradation by storing multiple aliquots Sample security Cost effectiveness Cost 3-Nov-14

12 Questionnaires 問卷 Child’s questionnaire Diet Physical activity
General health Moods and feelings Future development Microbiota Sleeping habit Respiratory health Parents’ questionnaire Physical health Social skills Infant diet Parents’ sleeping habit Socio-economic status 3-Nov-14

13 Physical assessment forms 體格檢查表
Height Weight Sitting height Head circumference Arm circumference Waist circumference Hip circumference Grip strength Blood pressure Body composition Pubertal stage 3-Nov-14

14 Schedule 時間表 Clinics open at Saturday & Sunday
Friday during summer holidays Public holidays 3-Nov-14

15 Preparation work before clinics
2 Registration 預約 3 Confirmation letter & stool pack 確認信&糞便樣本收集包裹 1 Invitation letter 邀請信 Preparation work before clinics 5 Reminder call 提醒備忘 4 Fieldwork documents (barcode labels, lab forms) 文書 (條形碼標籤,實驗室送檢表格) 3-Nov-14

16 Sample collection at clinics
1 Anthropometric measurements 體格檢查 Height Weight Head, arm , waist, hip circumference Blood pressure Pubertal stage Grip strength 3 Lung function test 肺功能測試 2 Blood taking 抽血 30 ml FEV1 FVC Sample collection at clinics 4 Sample collection 生物樣本 Stool Urine (9 ml) Saliva (2-5 ml) Hair (0.15g) Toenails (10) 5 Questionnaires 問卷 Child’s questionnaire Parent’s questionnaire Coupons $200 禮券 Examination report 檢查報告 3-Nov-14

17 On site procedures - anthropometric measurements
Bio-impedance analysis (BIA) 生物電阻分析 Blood pressure 血壓 3-Nov-14

18 On site procedures - blood taking
抽血 3-Nov-14

19 On site procedures - lung function
肺功能測試 3-Nov-14

20 On site procedures - sample collection
Stool sample 糞便樣本 Hair sample 頭髮樣本 3-Nov-14

21 Instructions for stool sample collection
Place all gel packs needed in your freezer for at least 12 hours Use the scoop on the cap to collect stool sample into the bottle. Put the stool collection bottle inside the biohazard ziplock bag and seal the bag. Put paper cup containing your stool specimen on top of the gel pack in the cooler bag. 3-Nov-14

22 Examination report 3-Nov-14

23 Sample processing and storage
Clinics: Original samples from participants 診所:原始樣本 1 Laboratory : Processed samples in aliquots 實驗室:處理、等分後樣本 2 Sample processing and storage Storage center: Cryotubes & cryoboxes in -80 oC freezer 存儲中心:凍存管儲存於-80度冷凍箱 3 3-Nov-14

24 (tubes in close contact with ice cubes/ ice packs)
Blood sample transport Biohazard Bag c Biohazard Bag a 1 x 5mL Clotted (Orange) 1 x 2mL Fluoride-oxalate 1 x 3mL EDTA Department of Chemical Pathology lab form Serum cryolabels Department of Hematology lab form Biohazard Bag b 4 x 3mL EDTA Whole blood cryolabels Blood spot general labels Buffy coat cryolabels Plasma cryolabels Biohazard Bag d 1x 5mL Clotted (Red) Chilled in ice during temporary storage Cooler Box PWH-2 (4ºC) (with few ice packs) Cooler Box PWH-3 (<4ºC, chilled) (tubes in close contact with ice cubes/ ice packs) Cooler Box PWH-1 (<25ºC) (with few ice packs)

25 Taxi from Clinic to PWH, arrive at around 12:00pm
(Biohazard bag a) Deliver to Department of Chemical Pathology, PWH (Sample reception, 3/F, New Clinical Building, PWH) (Biohazard bag c) Deliver to Department of Hematology, PWH (Room 34051, 1/F, Clinical and Sciences Building, PWH) Deliver to Department of Pediatrics, PWH (Room 903, 9/F, Li Ka Shing Medical Sciences Building, PWH) Deliver to Department of Chemical Pathology, PWH (Sample reception, 3/F, New Clinical Building, PWH) Confirm the sets of samples given to the PWH staff (Applies to all) Blood analysis and storage Further processing Blood analysis Blood analysis and storage

26 Saliva, urine and stool sample transport
Cooler Box (<25ºC) (with few ice packs) 1 x saliva (microbiome) sample Cryolabels for saliva Biohazard Bag e(1) Public transport from clinic to SPH-HKU Further processing at SPH lab 1 x stool sample Cryolabels for stool Biohazard Bag e(2) 1 x urine sample Cryolabels for urine SPH: School of Public Health

27 Public transport from clinic to
Hair and toenail sample transport Biohazard Bag f(1) Ziplock bags (<25ºC) 1 x hair (no roots) sample 1 x toenails sample Public transport from clinic to SPH-HKU Storage Biohazard Bag f(2) 1 x saliva (DNA kit), if applicable SPH: School of Public Health

28 Blood sample processing
RNA Whole blood spot card 0.5ml Clotted 5ml Insulin, hsCRP (1ml) Fluroride Oxalate 2ml EDTA 3ml ??? Hormones antibodies DNA Immortal cells Buffy coat 0.2ml + glycerol Hormone Clot factor 1 2 3 4 5 6 Serum 0.8ml Whole blood Plasma 0.25ml + trizol 0.75ml 4.8ml red cells + trizol LS 0.6ml 1.2ml buffy coat plasma Slow spin Fast spin RFT, LFT, lipids glucose hbA1c CBC 7 8 9

29 Barcode 條形碼 L: Laboratory sample BC : Birth Cohort 01: # of sample
13: year 2013 SE:sample type (serum) 100001: participant ID 01: # of sample L: Laboratory sample 3-Nov-14

30 Blood sample storage …. 血液樣本 SE1 (#1) SE2 (#1) SE3 (#1) SE4 (#1) SE5
Processing SE denotes sample type (Serum) Each participant has 5 aliquots of serum, 0.8ml each (SE1-SE5) Blood sample from several participants #1, #2…. …. SE1 (#2) SE2 (#2) SE3 (#2) SE4 (#2) SE5 (#2) Paste the corresponding cryolabel on the tube (vertical and not blocking the window)

31 First, scan Box A barcode
Select the inventory Login to the lab system https://labsystem.influenza.hk/ Select freezer First, scan Box A barcode Select corresponding boxes for the sample type (Check label description in SOP or on the label) Repeat the process for subsequent samples in the listed direction and press confirm when finish SE1 (#2) SE2 …. Scan barcode on cryolabel SE1 (#1) SE2 ….

32 Put the boxes , which contains the samples,
back into the -80 oC freezer

33 Saliva sample collection and processing
DNA kit DNA Transport at ~25 oC and store at 4 oC at SPH-PMB (Only for those without blood sample) Collection of saliva (DNA) sample Participant will Spit into Collection Funnel Remove Collection Funnel Add and mix the contents of the Preservative Ampoule with the collected saliva Saliva 1 ml Microbiome Transport at 4oC 2-5ml in a pot Step 1 Collection of saliva (Microbiome) sample Participant will Let saliva collected in the mouth for at least 1minute. Drool into the labeled collection pot (Yellow cap). Repeat multiple times until collecting 2-5ml. Step 2 Processing of In laboratory, Aliquot saliva from collection pot into 2 aliquots of saliva (1.0 ml each) Store the 2 aliquots of saliva sample in cryostorage tubes Store at -80 oC 唾液樣本 SPH: School of Public Health; PMB: Patrick Manson Bldg office

34 Transport and store at ~25 oC
Urine sample, hair and toenails collection and processing Step 1 Collection of ‘mid-stream’ urine sample In washroom, participant will Wash and dry hands Unscrew the lid of urine collection pot Pass small quantity of urine into toilet Pass urine into collection pot (Yellow cap) until about half full Step 2 Processing of urine sample In laboratory, Aliquot urine from collection pot into 3 aliquots of urine (about 1.0 ml each) Store the 3 aliquots of urine sample in cryostorage tubes Transport and store at ~25 oC SPH-PMB Toenails Chemical exposure Hair 0.15 g Urine 1/2 pot 1 ml Store at -80 oC Transport at 4oC Microbiome Collection of hair (Chemical exposure) Research assistant will Wear a pair of new gloves before collection Cut few pieces of hair (0.15g) from each participant using a pair of scissors Collection of toenails (Chemical exposure) At home, participant will Clip the toenails and place it inside the labeled nail collection bag and bring it to the clinic 尿液樣本 頭髮、趾甲樣本

35 Stool sample collection and processing
1/3 pot Stool sample collection and processing Step 1 Collection of stool sample within 16-hour before the visit Step 2 Packing of stool sample for delivery Step 3 Processing of stool sample Store at -80 oC Transport at 4oC 1 ml In laboratory, Use a pre-sterilized fork and transfer a small lump of stool (200ul mark) Store the 2 aliquots of stool sample in cryotubes 糞便樣本

36 “Children of 1997” Biobank Based on a large, population representative Chinese cohort First generation of Chinese children to grow up in a high-income setting with a social infrastructure similar to the Western countries Key developmental stage (16-18 years) What is special about our Biobank? Sample storage condition Fresh sample vs -80 ºC storage Effect of reagent Potential problems Regular quality test Combine processing site and storage center Samples from each participant stored in 2 locations Tablet for questionnaire Further improvements 3-Nov-14

37 Collaborators 3-Nov-14

38 Funding sources WYNG Foundation (for Biobank)
Health and Medical Research Fund General Research Fund Hong Kong Health Care and Promotion Fund Health and Health Services Research Fund Research Fund for the Control of Infectious Diseases The University of Hong Kong, University Research Council Strategic Theme of Public Health 3-Nov-14

39 Thank you ! For further information: Dr. Mary Schooling: cms1@hku.hk
Dr. Cindy Lin: 3-Nov-14


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