Tecan Group AG · SIX:TECN
Tecan — 3rd Tecan Symposium, Salzburg 2010
66 slides from Tecan's conference presentation, Oct 7, 2010
Slide 1: Molecular Testing and the Changing Face of Medicine
Molecular Testing and the Changing Face of Medicine A Quick Walk through the Park James K. Fleming, PhD, FACB Vice President and Director Department of Science and Technology Laboratory Corporation of America Elon, North Carolina USA 3rd Tecan Symposium, Salzburg October 7, 2010
Slide 2: A Sinuous Walk Through the Park
2 A Sinuous Walk Through the Park
Slide 3: 3
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Slide 4: The Beginning
4 The Beginning
Slide 5: History of Molecular Testing
5 History of Molecular Testing 1953 – Jim Watson and Francis Crick publish the structure of DNA (Nobel Prize 1962) 1957 – Kornberg identifies DNA polymerase which requires an initiation primer (Nobel Prize 1959)
Slide 6: History of Molecular Testing
6 History of Molecular Testing 1960 – Khorana discovers the Genetic Code (Noble Prize 1968) 1969 – Brock isolates Thermus aquaticus from a hot spring in Yellowstone National Park
Slide 7: History of Molecular Testing
7 History of Molecular Testing 1970 – DNA polymerase is isolated from E. coli 1971 – Kleppe and Khorana theorize that 2 primer system could lead to replication of a specific segment of DNA, but publish no data 1976 – DNA polymerase is isolated from T. aquaticus (Taq)
Slide 8: History of Molecular Testing
8 History of Molecular Testing 1977 – Sanger reports a method of sequencing DNA (Nobel Prize 1980)
Slide 9: History of Molecular Testing
9 History of Molecular Testing Between 1953 and 1980, these researchers set the stage for the next level of discovery by providing all the components needed to perform PCR (4 Nobel awards) Structure Primers Polymerase Replication Sequencing
Slide 10: History of Molecular Testing
10 History of Molecular Testing 1983 Kary Mullis at Cetus modified the amplification process by bracketing a desired sequence of the DNA strand with 2 primers using thermocycling Then used DNA polymerase to make infinite copies
Slide 11: History of Molecular Testing
11 History of Molecular Testing Problem: DNA polymerase was thermal labile 1985 – 1st PCR publication appears in Science 1986 Cetus files PCR patents Mullis uses Taq polymerase First forensic use of PCR for HLA‐DQA 1988 Perkin Elmer launches first automated thermal cycler Science prints
Slide 12: History of Molecular Testing
12 History of Molecular Testing 1989 – Star Trek: The Next Generation. The ships doctor is aging rapidly by a virus and is cured when her pre‐infection DNA isolated from a strand of her hair is found in her room. PCR hits mainstream media
Slide 13: History of Molecular Testing
13 History of Molecular Testing 1990‐ Fist forensic PCR kit for HLA‐DQA 1991– Roche secures world wide rights for PCR patents from Cetus for $300M US 1992 – Roche Molecular Diagnostics is founded Launch Amplicor HIV‐1 Monitor x‐US 1993 – Launch of the 1st commercial kit in US for Chlamydia tra
Slide 14: History of Molecular Testing
14 History of Molecular Testing Roche also launches the first multiplex Amplicor CT/NG PCR 2000 – Roche launches Linear Array CF test 2001 – Roche patents real time PCR devise 2001 ‐ 2010
Slide 15: Molecular Testing
Molecular Testing 15 Genetic Abnormalities Oncology Neurology Prenatal Coagulation Pharmacodiagnostics Tissue typing Endocrine Connective Tissue Hematology Renal Transfusion Medicine
Slide 16: Molecular Testing
16 Molecular Testing Infectious Organisms Bacterial • Mycobacteria Viral Rickettsial Fungi
Slide 17: Molecular Techniques
17 Molecular Techniques PCR Non‐PCR Amplification bDNA TMA SDA LiPA NASBA ISH Sequencing
Slide 18: Other Molecular Platforms
18 Other Molecular Platforms Microarray Laser Capture Microdissection Tissue Arrays Amplification coupled LC/MS Microfluidics Invader
Slide 19: Top Molecular Companies
19 Top Molecular Companies Abbott Cepheid Qiagen Agilent Dako Roche BD Genomic Health Siemens Beckman Coulter GenProbe Sysmex bioMerieux Genzyme Ventana Celera Ortho Plus 130 additional companies providing molecular diagnostics from AdnaGen, AG to Zila, Inc
Slide 20: The Impact on Medicine
20 The Impact on Medicine
Slide 21: HIV: The Beginning
21 HIV: The Beginning The 1st Culprits Chimpanzee SIVcpz HIV-1 Sooty Mangabey SIVsm HIV-2
Slide 22: HIV: The Beginning
22 HIV: The Beginning The 2nd Culprits Contaminated Needle Theory Hunter Theory
Slide 23: HIV: The Beginning
23 HIV: The Beginning Oral Polio Vaccine Theory Conspiracy Theory
Slide 24: HIV: The Beginning
24 HIV: The Beginning Earliest Evidence 1959 – plasma from adult male living the Congo 1960 – lymph node of adult female living in Congo 1969 – tissue of dead teenager living in St Louis 1976 – tissue sample Norwegian sailor 1979 – Gay men in US and Sweden and heterosexuals in Tanzania an
Slide 25: PCR and HIV
25 PCR and HIV June 1981 – Brief report in MMWR of unusual pmeumocystis carinii pneumonia among “5 gay, otherwise healthy men” from LA. PCP almost always seen in immunocompromised patients The 5 men had no significant medical history July 1981 – 45 more cases isolated in LA, SF, NY Some deve
Slide 26: PCR and HIV
26 PCR and HIV 1984 – 4000 cases in the US including • hemophiliacs • infants born to HIV infected mothers • Men who have sex with men • The disease now called AIDS French researchers discover a retrovirus NIH announces that a vaccine will be ready in 2 years
Slide 27: HIV: The Beginning
27 HIV: The Beginning Early developers of HIV tests were faced with an unmet clinical need for testing viral infectious diseases Need to detect low antibody levels Need early detection Need to follow viral concentration in host
Slide 28: PCR and HIV
28 PCR and HIV 1985 – 1st HIV antibody test became available (ELISA) 1986 – Switzerland begins testing blood products 1987 – ATZ (zidovudine) becomes 1st anti‐HIV drug at a dose of 100mg q4h ‐24 hours per day Western Blot testing begins 1988 – Deaths in US at 4,855
Slide 29: PCR and HIV
29 PCR and HIV 1989 – Deaths in US at 14,544 Reverse transcriptase inhibiters released 1992 – First rapid test for HIV antibody released Deaths in US at 23,411 1994 – HIV ability to mutate have made vaccine development difficult and risky 1995 – Roche launches HIV‐1 PCR quant
Slide 30: HIV and PCR
30 HIV and PCR 1996‐ Era of HAART begins (3 drugs recommended) Deaths in US at 21,399 Estimated 22M infected WW with 6.4M deaths 1999 – NIH pledges to develop a vaccine within the next 10 years HIV ‐1 ultra sensitive PCR test launched Deaths in US at 17,741
Slide 31: PCR and HIV
31 PCR and HIV 1999 – Monogram launches anti‐retroviral susceptibility testing Tropism for CCR5 or CXCR5 for CCR5 agonists Phenotypic and genotypic susceptibility • non‐nucleoside reverse transcriptase inhibitors • nucleoside reverse transcriptase inhibitors • protease inhibitors • Integrase •
Slide 32: PCR and HIV
32 PCR and HIV 2002 – Roche HIV‐1 v1.5 PCR screen for blood and blood products launched in Italy 2007 – 2.7 M have been infected in US and 2.0M have died HIV‐1 RT viral load launched by Roche and Abbott (Abbott claims greater detection of subtypes) 2010 – Roche releases v2 of HIV‐1 RT PCR with
Slide 33: HIV and Changes in Medicine
33 HIV and Changes in Medicine Several influencing factors PCR • Drug discovery • Patient diagnosis • Patient monitoring • Selective patient treatment Pharmaceuticals and HAART Education HAART has decreased death rate by 80% and increased life expectancy by 20 years
Slide 34: HIV and Changes in Medicine
34 HIV and Changes in Medicine In 1985 Median time of progression to AIDS – 9 to 10 y Median survival time – 9.2 months In 2010 Survival time has increased by 4 to 12 y
Slide 35: AIDS Diagnoses and Deaths of Adults and Adolescents
AIDS Diagnoses and Deaths of Adults and Adolescents with AIDS, 1985-2007— United States and 90- Dependent Areas Diagnoses =@Deaths (in “ £ & wu mo) mo} c © wv 7] “ °o = 122) po mo] rr f°] ) = 1985] 1986] 1987] 1988] 1989] 1990] 1991] 1992] 1993 | 1994] 1995] 1996] 1997] 1998] 1999] 2000] 2001| 2002]
Slide 36: Adults and Adolescents Living with an AIDS Diagnosis
Adults and Adolescents Living with an AIDS Diagnosis, by Sex, 1993-2007—United States and Dependent Areas NO ND o wo o (in thousands) rn) Females “a < 3° “ he Q a i) (} = = o o 1993] | 1995| 1996| 19971 1998] 1999| 2000 | 2001] 20021 2003| 2004] 2005 | 2006| 2007] Year of diagnosis Note. All display
Slide 37: Median Age at Death due to HIV Disease
Median Age at Death due to HIV Disease United States, 1987-2006 = - © ® Q — © ® D < e = a] ® = LJ LJ LJ LJ Ly q qT q ! i LJ LJ ! I T q i 1987 1988 1989 1990 1991 1992 1993 1994 1995 1996 1997 1998 1999 2000 2001 2002 2003 2004 2005 2006 Pc st3¥ice, ty : 2 (4 Note: For comparison with data for 1999 a
Slide 38: Estimated Numbers of AIDS Cases, Deaths, and Persons Living with AIDS
Estimated Numbers of AIDS Cases, Deaths, and Persons Living with AIDS, 1985-—2007—United States and Dependent Areas ” & 5 Oo {00} UG Ze © § nos oo AS Se OF (eo) fe} ra “| A Cases Deaths @ Prevalence i<—— 1993 definition implementation 500 450 400 + 350 10]0) + 250 Prevalence thousands) + 200 in ( 15
Slide 39: But Not Globally
39 But Not Globally 95% of new HIV infections arise in lower to middle income nations <1% of sexually active people of Africa’s urban population has been tested. Over 33M people are currently infected 31M adults 15.7 women (50%) 2.1M children
Slide 40: But Not Globally
40 But Not Globally 25M have died from AIDS since 1981 More than ½ of all those who are infected are less than 25 yo • Will likely die before they are 35 AIDS deaths in 2008 – 2M WW WHO estimates only 5.2M receiving treatment
Slide 41: Companion Diagnostics in 2010 Anti-retrovirals
Companion Diagnostics in 2010 41 Anti‐retrovirals NRTI NNRTI PI Integrase FI Zidovudine Nevirapine Saquinavir Raltegravir Enfuvirtide Didanosine Delavirdine Ritonavir Zalcitabine Efavirenz Indinavir Stavudine Etravirine Nelfinavir Lamivudine Atazanavir Abacavir Fosamprenavir Tenofovir Tipranavir 6 d
Slide 42: PCR and HIV
42 PCR and HIV Medical science can help It has the means and ability to increase the quality of life Global economics, governments and education remain barriers PCR has changed the practice of medicine for a pernicious and deleterious disease But its influence is limited.
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Slide 44: Molecular Testing and Controversy
44 Molecular Testing and Controversy The Case of MRSA 2007 CMS issued a directive that the agency would no longer pay for complications arising from hospital acquired conditions if that condition was not present at time of admission. Hospitals immediately sought a molecular solution for methicil
Slide 45: Molecular Testing and Controversy
45 Molecular Testing and Controversy CMS decided to delay its rule MRSA molecular screening interest precipitously dropped in US In preference to Remel’s selective media Lower cost Doesn’t eliminate the need for rapid detection Does set up a mechanism for substandard treatment and patient cr
Slide 46: Molecular Testing and Controversy
46 Molecular Testing and Controversy Direct to Consumer Testing Posed scientific, ethical, and moral dilemmas for the US Can have our entire genome scanned for $1000‐ $3000 Is there a benefit? Or should we selectively genotype specific disease causing regions of our DNA The genotype is ac
Slide 47: Molecular Testing and Controversy
47 Molecular Testing and Controversy Interpretation may vary from one company to another Is it beneficial to know if one has a predisposition to heart disease or diabetes – much debate • Could lead to undue stress, anxiety, needless decisions Genetic information should be considered with other
Slide 48: Molecular Testing and Controversy
48 Molecular Testing and Controversy Part of problem • Genetic information frequently incomplete Unlike Huntington’s or CF Heart Disease, Diabetes, Cancer, • Genetics are complex No single genes May be tens of genes that have correlative mutations yet to be discovered Interpretations provide ris
Slide 49: Molecular Testing and Controversy
49 Molecular Testing and Controversy Negative Test does not mean your free of risk or disease Environmental Factors Population bias All pose caveats and limitations that change the clinical significance of the mutations detected
Slide 50: Who Owns Our Genes
50 Who Owns Our Genes ~25,000 protein encoding genes 20% of genes are patented US Patent Office has received more that 3M gene Round Worm applications over past 20 ~20,000 genes years 40,00 patents held on gene segments
Slide 51: Who Owns Our Genes
51 Who Owns Our Genes 1906 – Adrenalin was patented Patent was challenged and US Supreme Court ruled that the substance was “isolated and purified” and that constituted an invention regardless of it’s source 1923 – Insulin was patented 1980 – Diamond v Chakrabarty GE engineer who developed a
Slide 52: Who Owns Our Genes
52 Who Owns Our Genes Patent challenge resulted in the US Supreme Court ruling that GE patent was valid • The organism was man made therefore it was an invention Supreme Court ruling led to liberal interpretation of the rulings by the Patent Office and began applying the rulings to genetic materi
Slide 53: Who Owns Our Genes
53 Who Owns Our Genes March 29, 2010 ACLU v Myriad Genetics • Myriad holds patents for several genes in the BRCA family • Mutations of BRCA incur susceptibility to breast and ovarian cancer • Myriad also has property rights to the diagnostic test which is sold for $3000 US
Slide 54: Who Owns Our Genes
54 Who Owns Our Genes ACLU argued that the gene patents stifled research, innovation and limited testing options Attorney Generals Office argued that the Patent Office was interpreting court rulings and Congressional law Federal Court invalidated 7 Myriad IP patent for BRCA Myriad vowed to appea
Slide 55: Cartoon on breast cancer genes
Whey re our breast Cancer % genes —we idenhfied then. —¢ It's kind of you fe let uS have the disease LabCorp
Slide 56: The Prospects
56 The Prospects
Slide 57: The Future
57 The Future Do gene patents stifle research? Do companies sit on patents that could led to new therapies in lieu of working on blockbuster genes that could result in lucrative returns Or will loss of gene patents disincentivize companies from active pharmacodiagnostic research Will invalida
Slide 58: The Future
58 The Future Will other countries see court action due to less conscientious molecular laboratories Will professional organizations rally to the cause and self police through stricter laboratory operating guidelines and certification programs
Slide 59: The Future
59 The Future Will governmental agencies pass laws governing molecular diagnostic practices for the protection of the consumer Many states in US have laws banning the use of genetic testing to determine insurability
Slide 60: The Near Future from LabCorp's Perspective
60 The Near Future from LabCorp’s Perspective Increased decentralization of molecular testing Molecular testing equipment is becoming more black box No longer a central laboratory test • CT/NG • HPV • HIV • HBV • HCV
Slide 61: The Near Future from LabCorp's Perspective
61 The Near Future from LabCorp’s Perspective Alternate Sample Types
Slide 62: The Near Future from LabCorp's Perspective
62 The Near Future from LabCorp’s Perspective Microfluidics PCR coupled technology Multiplexing for Infectious Organisms
Slide 63: The Near Future from LabCorp's Perspective
63 The Near Future from LabCorp’s Perspective Single cell PCR Circulating Tumor Cells Increased microarray applications for clinical trials R&D diagnostics
Slide 64: The Future from LabCorp's Perspective
64 The Future from LabCorp’s Perspective Changing role for molecular biologists • Directors rather than bench technologists Expanded role of DNA evidence in pretrial and post conviction cases
Slide 65: The Future ?
65 The Future ?
Slide 66: The Break!
66 The Break!