TM

COLOTECT

COLOTECT TM

Introduction

International Business Development Department

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COLOTECT
Epidemiology of colorectal cancer

Mortality
- Almost 861,000 deaths every year

Incidence
- 1.8 million new cases occur every year

Risk
- Age

- Personal or family history Colostomy after Colorectal cancer surgery

3,000,000
Preventable
- Screening is an important step in
finding early stages of CRC. Patients with colostomy worldwide [1].

Winawer SJ, Zauber AG, Ho MN, et al. Prevention of colorectal cancer by colonoscopic polypectomy. The National Polyp Study Workgroup [Internet]. The New England Journal of Medicine (US); 1993 Dec 30 [cited 2023 Apr 22]. Available from:
https://europepmc.org/article/med/8247072
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Cost effectiveness compared to no screening

CRC Development

Stage Stage
CRC Staging Early Stage Stage I Stage II
III IV

5-year Survival Rate 90% 71% 14%

Localized Stage Spread to surrounding Spread to distant parts

tissue/organ

REFERENCE:
1. N Engl J Med, 1993, 329(27): 1977-1981.
2. Available: https://gutscharity.org.uk/2019/05/growing-numbers-of-under-50s-diagnosed-with-bowel-cancer-in-the-uk/
3. Prz Gastroenterol. 2019; 14(2): 89–103.
4. Available at https://www.physiciansweekly.com/colorectal-cancer-screening-cost-effectiveness/
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Guidelines for Colorectal Cancer Screening

Recommendation
Who Should Be Screened for Colorectal Cancer? How frequently the screening should be done?

• Asymptomatic individuals at age 50 to 74 are • Every 2 years

recommended for the iFOBT population screening
• Asymptomatic individuals aged 45 to 49 years can
request the screening

Colonoscopy Stool DNA testing

• Every 5-10 years • Every 3 years

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COLOTECT
Current Screening Methods

Colonoscopy FOBT DNA-Methylation

• Recommended once every 5 to 10 years • Recommended once every 1 to 3 years • Recommended once every 3 years
Pros: Pros: Pros:
• Golden Standard for CRC Dx • Non-invasive • Non-invasive
• High sensitivity and specificity • Less expensive • More reliable: Sensitivity as high as 87% to 92%
• Suspicious polyps can be removed during the • Sample can be collected at home or clinic and in precancer and early stage CRC
process sent to the lab
• Stool DNA (sDNA) is continuously shed into stool
Cons: Cons: it will detect cancer even without an actively
bleeding lesion
• Invasive • Limited sensitivity in precancer (73% sensitivity)
• Sample can be collected at home or clinic and
• Requires dietary restriction and at least one day • Sample can be collected at home or clinic and sent to the lab
for preparation sent to the lab
• Must be performed at the hospital or clinic with • False negative for CRC patients without actively Cons:
an an experienced gastroenterologist in site bleeding lesions
• More expensive than FIT (but cheaper than
colonoscopy)

Why we screen for some cancers and not others [Internet]. American Cancer Society (US); 2019 Sep 9 [cited 2023 April 22]. Available from: https://www.cancer.org/latest-news/why-we-screen-for-some-cancers-and-not-others.html
Castells A, Quintero E. Programmatic Screening for Colorectal Cancer: The COLONPREV Study [Internet]. Springer Science+Business Media (Germany); 2015 Mar [cited 2023 April 22]. Available at https://link.springer.com/article/10.1007/s10620-014-3446-2
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 FIT: low sensitivity

• Fecal immunochemical test (FIT) is used for detecting invisible hemoglobin in red blood cells
on the surface of stool.
• The principle of FIT is antigen-antibody reaction which has specificity to human hemoglobin in
lower digestive tract .
[1]

• Cancer and precancer may bleed intermittently . [2]

24% Sensitivity for

advanced adenoma [3] 79% Sensitivity for
CRC [3]

Source: [1]https://www.eiken.co.jp/en/ourfields/gastroenterology/whatisfit/
[2]Exact Science, Cologuard Stool DNA testing for Colorectal cancer screening (physician orientation), p3
[3]Bénard F, Barkun A N, Martel M, et al. Systematic review of colorectal cancer screening guidelines for average-risk adults:
Summarizing the current global recommendations[J]. World journal of gastroenterology, 2018, 24(1): 124..

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Limited resources of Colonoscopy
Number of Endoscopists per Million People
300

250
250

200

150

100

50
22 17
0 6 5 4 4 2
0

Countries like China, Malaysia, Vietnam, India, Thailand, Philippines, Indonesia,

have very limited resources of Colonoscopy.
Number of endoscopist in local country remains to be investigated.
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COLOTECT
COLOTECT Characteristics

Effective
Precise Sensitivity as high as 87%
Specificity as high as 93% • Adenoma polyps over 1 cm
• Serrated adenoma
• Villous adenoma

Convenient
Easy, painless and private sample collection,
without special preparation

Simple Workflow
Uses commonly used PCR platforms, detects all Reliable
targets in one well, enabling quick processing and 3 biomarkers and 1 internal control
reporting

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Colotech VS FIT sensitivity
Sensitivity of COLOTECTTM and FIT(OC FIT-CHEK) [1]
87,60% 87,90%
90,00%
74% 70%
80,00%
70,00%
46,20%
60,00%
50,00%
40,00%
24%
30,00%
20,00%
10,00%
0,00%
All CRC Stage I & II Precacerous Lesion

COLOTECT FIT(OC FIT-CHEK）

Precancerous lesion includes: advanced adenoma, serrated lesions and other lesions with high-grade intraepithelial neoplasia

COLOTECTTM Fecal immunochemical test

Detects methylated DNA Detects blood(hemoglobin)
DNA biomarkers are continuously shed Cancer and precancer may bleed intermittently[2]
into stool
3 biomarkers 1 biomarkers
Source:[1] COLOTECT Retrospective multicenter clinical effectiveness study on 329 clinical specimens
[2] Exact Science, Cologuard Stool DNA testing for CRC Screening for physicians, 2017.
© 2021 BGI Page
All rights 3
reserved.
 Colotech VS FIT PPV
• Fecal occult blood testing as a screening method for colorectal cancer leads to a high rate of
positivity, but a low rate of abnormalities diagnosed after further colonoscopy in those who are
tested positive.

Screening method Positive rate Abnormality rate by

colonoscopy（Polyp, adenoma,
cancer）
Fecal occult blood test 14.7% 31.4%
The TARGET-C trial’s
screening data [1]

Stool DNA test 5.4% 68.83%

BGI COLOTECT - 260,000 participants’
screening data

Note: The positive rate of FIT test and abnormal rate by colonoscopy in Colorectal cancer screening is variable. In
2022, a paper published in Lancet shows that the positive rate of FIT in Colorectal cancer screening is 20.1%, and the
abnormal rate is 17.5%.

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COLOTECT
Comparison Among CRC Tests
BGI
Cologuard® Epiprocolon® EarlyTect ColoScape Fecal occult blood FIT* Fecal gFOBT* COLOTECT

Detection Multiplex fluorescent PCR

qPCR qPCR Xenonucleic acid and qPCR Immunoassay Chemical method Multiplex fluorescent PCR
Technology + immunoassay

Adenoma <17% (early)

- - 66.6% 27%-29% 11%-25% 42%
Sensitivity <42% (advanced)

Bowel Cancer
92-94% 68.2-72.2% 87% - 92.3% 88.9% 71%-75% 33%-75% 87%
Sensitivity [1]

Specificity 87-89% 80% 90.2- 95.2% w/FFPE up to 96% 94% 98% 93%

PCR
FIT+PCR PCR PCR PCR
Biomarker (APC, BRAF, Hemoglobin Hemoglobin
(KRAS, NDRG4, BMP3) (Septin 9) (SDC2) (SDC2, ADHFE1, PPP2R5C)
CTNNB, KRAS)

Stool *test can be

Sample Stool * Blood* Stool Blood Stool Stool
performed locally

Sample
Manual Manual Manual Manual N/A N/A Automatic DNA extraction
Processing

FDA/CE IVD, NCCN

Certification recommended, CMS FDA/CE IVD MSFD (Korea) CE IVD N/A N/A CE IVD
reimbursed

*Gies A, Cuk K, Schrotz-King P, Brenner H. Direct comparison of diagnostic performance of 9 quantitative fecal immunochemical tests for colorectal cancer screening [Internet]. American Gastroenterological Association (US). 2018 Jan [cited 2023 Apr 22]. Available from:
https://pubmed.ncbi.nlm.nih.gov/28958859/

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COLOTECT
COLOTECT Technology Justification
DNA Methylation in Tumorigenesis sDNA as a molecular marker
DNA methylation changes have been recognized as one of the • Epithelial cells are constantly shedded in stool
most common molecular alterations in human CRC
• sDNA is easy to extract, and quite stable
• Enables a higher sensitivity than plasma

Plasma Marker vascular invasion

Distant
? metastasis

Stool Markers (exfoliation)

Large I II III IV
Pre-cancer Cancer

1
Relationship Between DNA Methylation and Tumorigenesis Molecular Markers Release from Colorectal Neoplasms into Target Media

Ahlquist DA, Taylor WR, Mahoney DW, Zou H, Domanico M, Thibodeau SN, Boardman LA, Berger BM, Lidgard GP. The stool DNA test is more accurate than the plasma septin 9 test in detecting colorectal neoplasia [Internet]. Clinical Gastroenterology and Hepatology
© 2021 BGI All rights reserved.
(US); 2012 Mar [cited 2023 Apr 22]. Available from: https://pubmed.ncbi.nlm.nih.gov/22019796/
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COLOTECT
How is Methylation Detected?
Bisulfite conversion process
• Unmethylated Cytosine (C) is converted to Uracil (U)
• Methylated Cytosines are protected from this conversion

Relationship between DNA Methylation and Tumorigenesis

Available from: https://www.diagenode.com/en/categories/bisulfite-conversion

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COLOTECT
COLOTECT Markers

[1]
SDC2 [2]
ADHFE1

[3]
PPP2R5C BGI Data
1 Han YD, Oh TJ, Chung TH, Jang HW, Kim YN, An S, Kim NK. Early detection of colorectal cancer based on presence of methylated syndecan-2 (SDC2) in stool DNA [Internet]. Clinical Epigenetics Society (Germany); 2019 Mar [cited 2023 Apr 22]. Available from:
https://pubmed.ncbi.nlm.nih.gov/30876480/
2 Fan J, Li J, Guo S, Tao C, Zhang H, Wang W, Zhang Y, Zhang D, Ding S, Zeng C. Genome-wide DNA methylation profiles of low- and high-grade adenoma reveals potential biomarkers for early detection of colorectal carcinoma [Internet]. Clinical Epigenetics Society
(Germany); 2020 Apr 21 [cited 2023 Apr 22]. Available from: https://pubmed.ncbi.nlm.nih.gov/32317010/
3 Xie H, Mahoney DW, Foote PH, Burger KN, Doering KA, Taylor WR, Then SS, Cao X, McGlinch M, Berger CK, Wu TT, Hubbard JM, Allawi HT, Kaiser MW, Lidgard GP, Ahlquist DA, Kisiel JB. Novel Methylated DNA Markers in the Surveillance of Colorectal Cancer Recurrence © 2021 BGI All rights reserved.
[Internet]. American Association for Cancer Research (US); 2021 Jan [cited 2023 Apr 22]. Available from: https://clincancerres.aacrjournals.org/content/27/1/141
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COLOTECT
COLOTECT Markers

COLOTECT PRODUCT A

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COLOTECT
COLOTECT Lab Workflow
High recommend!
• recommend qPCR machine
• automation processor
End User Slan 96S LineGene QuantStudio 5
9600 Plus

Sample collection Sample delivery DNA extraction Bisulfite qPCR Test report
conversion

Clinical Service Provider

Purifier HT Automatic Processor Purifier HT Automatic Processor

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COLOTECT
COLOTECT Experiment Workflow
TAT: ~2 days
Key Steps of Colotect Experiment Workflow
Without automation With automation
machine machine Throughputs: 5-10 plates of Samples/Week,
Room Steps Manual Manual Day
Duration Duration on 465-930 samples/week;
operation operation
on Device Device
time time
Reagent Reagent Preparation &
Technician: 2-3 (without automation), 1-2 (with
30min 0 1h 0
preparation Aliquot* automation);
Stool sample pretreatment 30min 0 30min 0 Responsibilities:
1ml supernatant fluid 1st • Task list arrangement, stool DNA extraction,
DNA
transfer into Deep-well 30min 0 30min 0 Day DNA bisulfite conversion, QPCR detection
Extraction
plate
Lysis 1h 0 1h 0 and Data analysis;
Extraction with Genfine 1.5h 0 10min 50min
• Fill in daily laboratory records;
DNA Bisulfite conversion 10min 2.8h 10min 2.8h
bisulfite • Reagent and consumables preparation.
Purification 1.5h 0 10min 1h
conversion

qPCR qPCR amplification 2nd * Reagent Preparation could be in parallel with sample
20min 2h 20min 2h
detection (not including reagent prep) Day pretreatment.
Data
Analysis 20min 0 20min 0
analysis © 2021 BGI All rights reserved.
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COLOTECT
COLOTECT Kits

Reagent Description

Mix I PCR probes and primers

DNA polymerase, reaction

Mix II
buffer and dNTPs

Positive control Methylated control DNA

Negative control Unmethylated control DNA

① ④
Stool Sample
Methylation • 50 tests per kit
Detection
Collection
(qPCR) • Shelf life: 12 months

② ③ Contents 50 tests per kit

sDNA Bisulfite
Extraction Conversion Shelf-life 12 months

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COLOTECT
COLOTECT Collection Kits

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COLOTECT
COLOTECT Lab Workflow
Colotect Key Equipment List
Equipment List Description Intended use
Bio-safety cabinet Airtech BSC- Sample Preparation
1304ⅡA2
Water Bath DK-8B Sample incubator
Floor-standing 50ml tube/96 well Sample Centrifuge
Centrifuge plate
Nucleic Acid Purifier HT Automation for Stool
Extraction DNA extraction &
Machine bisulfite conversion
PCR Thermal Bioer TC-96/G/H(b)C bisulfite conversion
Cycler
Real time thermal SLAN-96S/ABI Methylation detection
cycler system Quanstudio5//LineGene • Reagent preparation room
9600 Plus • Extraction room
• Bisulfite conversion room
Digital Dry Baths Labnet Magnetic beads drying
• qPCR detection room.
(physically independent without direct
communication of air.)

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COLOTECT
COLOTECT Lab Solution qPCR Platform

Slan 96S

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COLOTECT
COLOTECT Lab Solution qPCR Platform

LineGene
9600 Plus

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COLOTECT
COLOTECT Lab Solution qPCR Platform

QuantStudio 5

Note: As QS5 curve is prone to instability and the threshold line is low, it is easy to misread Ct value
of curve. It is necessary to manually determine whether the Ct value is valid.
You are advised to determine the Amp Status in the exported data table
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COLOTECT
COLOTECT Clinical Sample Performance
SLAN-96S Applied Biosystems 7500
 Limit of Detection (LOD): Under the input amount of 20ng, the  Limit of Detection (LOD): Under the input amount of 20ng,
sample with a 2% methylation rate can meet 95% detection. the sample with a 2% methylation rate can meet 95%
 Diagnostic specificity/sensitivity: Fifty-one clinical samples was detection.
be used to test the diagnostic specificity/sensitivity in different  Diagnostic specificity/sensitivity: Fifty-one clinical samples
mechines. The diagnostic sensitivity was 84.21% and the was be used to test the diagnostic specificity/sensitivity in
diagnostic specificity was 93.33%. different mechines. The diagnostic sensitivity was 89.47%
and the diagnostic specificity was 93.55%.

LineGene 9600 Plus/Applied Biosystems

QuantStudio 5/Roche Lightcycler 480 II
Limit of Detection (LOD): Under the input amount
of 30ng, the sample with a 2% methylation rate can
meet 95% detection at LineGene 9600 Plus. Under
the input amount of 20ng, the sample with a 5%
methylation rate can meet 95% detection atApplied
Biosystems QuantStudio 5/Roche Lightcycler 480 II.

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Demo Report

Positive
It is recommended that a positive ColoTect screen be clinically correlated
and followed-up with a structural examination of the colon such as
diagnostic colonoscopy.

Negative
A negative result indicates a low likelihood that a colorectal cancer(CRC) or
an advanced adenoma(adenomatous polyps with more advanced pre-
malignant features) is present.
The chance that a person with or has an advanced adenoma is less.

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COLOTECT
COLOTECT Public Health Engagement
Under the leadership of the Municipal Party Committee and the Municipal Government, the Health Commission organized
efficiently and actively participated in 15 regions working with BGI. 120,000 people eagerly signed up for the test (the
reservation had to be closed due to the limited number of places), and 101,216 people completed the test within one
month.

Economics evaluation of colorectal cancer screening

Overall input Overall screened number (thousands) 101,216
Individual screened cost (RMB) 210
Direct medical cost (M RMB) 10.14
Overall input (M RMB) 31.4
Overall output (expense Number of Colorectal cancer: 108 2907
saved) advanced lesions Precancerous lesions and
expected to be adenomas: 1249
detected Polyps: 1550
Avoided cases of adenoma/polyp progression 240
to cancer
Direct and indirect screening benefits (M 345.93
RMB)
Cost benefit ratio 1:11 (345.93/31.4)

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COLOTECT
COLOTECT Testing Experience in China

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Thank you

30
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