PGx – Personalised Genetic Solutions

1. PGx – Personalised Genetic Solutions AceProbe Technologies (India) Pvt. Ltd., B-8, Namdhari Chambers, 9/54, D.B. Gupta Road, Karol Bagh, New Delhi-110005 Phone: +91-11-45098118 Fax: +91-11-23552378 Email: info@aceprobe.com; Website: www.aceprobe.com

2. Road less travelled! AceProbe - A Company with a difference Probing Molecules for Life Risk of disease and response to treatment varies from person to person. This is due to variation in human genetic coding, interactions between one’s genes and environment over a lifetime and the unique signature of the immune system. Defining the scope and nature of human biological variation allows the targeting of medical treatments to those most likely to benefit. Such treatments may include drugs or cell therapies tailored to a patient’s history, genes and immunology. AceProbe has evolved with a team of Biotech professionals and operates in the niche are of genomic that enable our partners to efficiently design and perform experiments, monitor progress, receive and interpret results,and making the adoption of technology in atrivial manner. Our comprehensiveTechnology Platform and System offeringsallow access to a multitude of clinical and research data, key analysis modules for both genotype and expression information, and database archiving and processing capabilities. Flexible platform and custom service features for a medium-density, ultra-high-throughput, customizable format make all its offerings unique with unmatched credentials and offers predictive outcomes for patients and delivering a personalized approach through genetics.

3. MISSION AceProbe®has been diligently working towards its mission to develop genetic tests that will become the model for comprehensive disease detection and informed treatment, ushering in a new era of predictive, preventative and affordable health care. We are dedicated to supporting healthcare providers with all the tools they need to go genetic and is helping lead the way in genetic testing to promote early disease detection and to help physicians achieve optimal dosing of critical medications. AceProbe® solutions include customized, validated assays and panels to support Cardiology, Oncology, Infectious Diseases &Nutrition.

4. FDA warns that poor metabolizers of PLAVIX™ are at increased risk for adverse cardiovascular events How can genetic variants affect Plavix™ effectiveness? CYP450 2C19 what is it? Plavix (clopidogrel) is activated in the body by the CYP4502C19 enzyme and works as an anticoagulant by inhibiting the receptor responsible for physiological activation of the platelet. The Plavix Mutation Panel Test analyzes a patient’s DNA for variations in the CYP4502C19 gene. In 2009 the Federal Drug Administration approved wording on the drug label encouraging genetic testing for DNA variants based on the following risks: * Compared to other patients, individuals with these variants are noted to have an approximate 30% de-crease in active metabolite in plasma * Approximately 30% of the US population will carry at least one variant allele at the CYP2C19 locus . * Patients who have at least one CYP2C19 variant can experience ad-verse reactions including serious or life-threatening vascular events when given the usual Plavix dose. Compared to other individuals, patients with these variants have an approximate 50% increase in risk of stroke, myocardial infarction or death. * Compared to other individuals with stent placements, patients with these variants have an approximate 3 fold increase risk of stent thrombosis. SPECIMEN SUBMISSION REQUIREMENTS * Collect 5 ml blood from adult in an EDTA anticoagulant tube * Store-refrigerate blood sample * Ship under 2-8 degree * Receipt by the laboratory within 24 hours of collection *Turn around time: 03 working days AceProbe Genetics Laboratories provides pick up services for your area, and specimens from offices outside the pick up area and reports can be mailed to the laboratory. On March 12, 2010, the FDA released a new Boxed Warning alerting healthcare professionals that tests are available to assess a patient’s genotype to determine if they are a poor metabolizer of PLAVIX™. * US Food and Drug Administration. FDA drug safety communication: reduced effectiveness of Plavix (clopidogrel) in patients who are poor metabolizers of the drug [safety announcement]. 2010 Mar 12 . * Mega JL, Close SL, Wiviott SD, Shen L, Hockett RD, Brandt JT, Walker JR, Antman EM, Marias W, Braunwald E, et al. Cytochrome P-450 polymorphisms and response to clopidogrel. N Engl J Med. 2009;360:354-62.

5. CYTOCHROME CYP 450 2C 19 determines Patient PLAVIX™ (Clopidogrel) Metabolism Before You Prescribe! METHODOLOGY AceProbe™ offers unparalleled detection rates using a Mass-Spectrometry assay for Cytochrome P-450 2C19 DNA by multiplexed PCR on MALDI-TOF MS platform. Mutations detected include : *2 c.681G>A, *3 c.636G>A, *4 c. 1A>G, *5 c.1297C>T & *(17) (-806C>T). CYP 2C19*17 Beyond the poor metabolizer alleles mentioned above, a recently identified CYP2C19 allelic variant (*17) (–3402C>T) has been associated with increased transcription of the CYP2C19 gene and with ultra-rapid metabolism of CYP2C19 substrates.9,11 CYP2C19*17 carriers appear to have an enhanced response to clopidogrel and may be at increased risk for bleeding.

6. Warfarin Sensitivity DNA Test CYP2C9: sets the rate for accumulation and elimination of Warfarin in the patient’s body. VKORC1: sets the effective concentration of Warfarin in the individual patient, and is associated with specific Warfarin maintenance dosing. Warfarinactivity is determined partially by genetic factors. The American Food and Drug Administration "highlights the opportunity for healthcare providers to use genetic tests to improve their initial estimate of what is a reasonable Warfarin dose for individual patients".[ Polymorphisms in two genes (VKORC1 and CYP2C9) are particularly important. VKORC1 polymorphisms explain 30% of the dose variation between patients:particular mutations make VKORC1 less susceptible to suppression by Warfarin. CYP2C9 polymorphisms explain 10% of the dose variation between patients,mainly among Caucasian patients as these variants are rare in African American and most Asian populations.These CYP2C9 polymorphisms do not influence time to effective INR as opposed to VKORC1, but does shorten the time to INR >4. Indications for Molecular Testing • Patients initiating Warfarin therapy • Patients with a history of unstable Warfarin dosing • Patients at risk for bleeding events while receiving Warfarin therapy Testing Methodology The DIAGNA-ACE ® Warfarin Sensitivity Molecular Assay is a diagnostic test for the detection of common allelic variants in cytochrome P450 2C9 and Vitamin K epoxidereductase complex subunit-1 (VKORC1) genes [including CYP2C9*1, CYP2C9*2 (3608C>T), CYP2C9*3 (42614A>C) and VKORC1 -1639G>A] which are key genetic factors contributing to individual variation in response to Coumarin-based anticoagulants. (AceProbe™ offers unparalleled detection rates using a Mass-Spectrometry assay for Cytochrome P-450 2C9 and VKORC1 by multiplexed PCR on MALDI-TOF MS platform.)

7. Peace of mind through genetic testing By administering a painless genetic test, your doctor will discover if you have genetic variations known to inhibit warfarin’s effectiveness. Acquiring this information is a huge step toward treatment that is safer, more effective and less costly in the long run. Responses to Warfarin treatment differ among patients. Factors such as age, weight and sex account for 10–20% of the differences. Genetic variations, however, account for up to 40%.3 So, by combining genetic testing with other considerations, your doctor can potentially address up to 60% of the factors that would cause you to respond negatively to Warfarin. That means getting you on a safe and stable dose quicker. And that means peace of mind—something you can’t express in percentages. Mind—something you can’t express in percentages. Worried about Warfarin? Ask your doctor about testing today. And start your Warfarin treatment with assurance instead of anxiety. Specimen Requirements Peripheral Blood—5 ml lavender-top (EDTA) tube. Invert several times to mix blood. Do not freeze; forward promptly at ambient temperature to the AceProbe® Lab or Collection Centre near to you. References Food and Drug Administration. New labeling information for Coumadin. Approved 1/22/2010. Anderson JL, Horne BD, Stevens SM et al. Randomized trial of genotype-guided versus standard warfarin dosing in patients initiating oral anticoagulation. Circulation 2007;116:2563-70. The International Warfarin Pharmacogenetics Consortium. Estimation of the warfarin dose with clinical and pharmacogenetic data. N Engl J Med2009;360:753-64. Gage B, Eby C, Johnson J, Deych E et al. Use of Pharmacogenetic and Clinical Factors to Predict the Therapeutic Dose of Warfarin.  Clinical Pharmacology & Therapeutics 2008: 84, 326–331 Sconce EA et al.  The impact of CYP2C9 and VKORC1 genetic polymorphism and patients characteristics upon warfarin dose requirements: proposal for a new dosing regimen.  Blood 2005;106(7):2329-33. Human Cytochrome P450 (CYP) Allele Nomenclature Committee; Warfarin Genotyping Reduces Hospitalization Rates, Including Those Due to Bleeding or Thomboembolism Based on a study by the Medco Research Institute™ and Mayo Clinic, presented at the 2010 Annual Scientific Session of the American College of Cardiology; Thomas P. Moyer et al. Warfarin Sensitivity Genotyping: A Review of the Literature and Summary of Patient Experience Mayo Clin Proc. • December 2009;84(12):1079-1094 Lee CR, Goldstein JA, Pieper JA. Cytochrome P450 2C9 polymorphisms: a comprehensive review of the in-vitro and human data. Pharmacogenetics 2002; 12:251-263 VKORC1 Genotype (up to 25%) Unknown Factors (at least 40%) CYP 2C 9 Genotype (up to 15%) 45 – 60 % AGE, Sex, Weight (at least 10-20%)

8. References •Lynch TJ, Bell DW, SordellaR, GurubhagavatulaS, OkimotoRA, BranniganBW, Harris PL, HaserlatSM, SupkoJG, HaluskaFG, Louis DN, ChristianiDC, SettlemanJ, HaberDA. Activating mutations in the epidermal growth factor receptor underlying responsiveness of non-small-cell lung cancer to gefitinib. N EnglJ Med. 2004 May 20;350(21):2129-39. •PaezJG, JännePA, Lee JC, Tracy S, GreulichH, Gabriel S, Herman P, Kaye FJ, LindemanN, BoggonTJ, Naoki K, Sasaki H, FujiiY, Eck MJ, Sellers WR, Johnson BE, MeyersonM. EGFR mutations in lung cancer: correlation with clinical response to gefitinibtherapy. Science. 2004 Jun 4;304(5676):1497-500. •Sharma SV, Bell DW, SettlemanJ, HaberDA. Epidermal growth factor receptor mutations in lung cancer. Nat Rev Cancer. 2007 Mar;7(3):169-81. •MokT, Wu, YL, ThongprasertS, Yang CH, ChuD, SaijoN, JiangH, Watkins C, Armour A, Fukuoka M. Phase III, randomised, open-label, first-line study of gefitinibvscarboplatin/ paclitaxelin clinically selected patients with advanced nonsmall-cell lung cancer (IPASS). Oral Presentation. ESMO 2008. •Kim ES, Hirsh V, MokT, SocinskiMA, GervaisR, Wu YL, Li LY, Watkins CL, Sellers MV, Lowe ES, Sun Y, LiaoML, OsterlindK, ReckM, Armour AA, Shepherd FA, LippmanSM, DouillardJY. Gefitinibversusdocetaxelin previously treated non-small-cell lung cancer (INTEREST): a randomised phase III trial. Lancet. 2008 Nov 22;372(9652):1809-18. •Kimura H, Fujiwara Y, SoneT, et al: High sensitivity detection of epidermal growth factorreceptor mutations in the pleural effusion of non-small cell lung cancer patients. Cancer Sci97:642-648, 2006 • EGFR Role in NSCLC ( Non Small Cell Lung Cancer) • Epidermal Growth Factor Receptor (EGFR) is overexpressed in various solid tumors including lung carcinoma • Activation of EGFR by ligand binding or mutation stimulates cellular tumor growth and proliferation. • Excessive activation of EGFR is associated with advanced stages of cancer and a poor prognosis. • Targeted Molecular Therapy • EGFR is a leading target for molecular based therapy in NSCLC. • Inhibition of EGFR pathway can impair cell proliferation and tumor growth. • Tyrosine kinase inhibitors (TKIs) selectively inhibit intracellular EGFR activity. • EGFR Mutations and Clinical findings • Studies suggest that patients with somatic mutations in the EGFR gene derive the greatest benefit from TKI treatments. • EGFR mutations have been reported in 10-20 percent of all NSCLC patients. • EGFR mutations have been reported in 75-90% of patients who clinically respond to EGFR TKIs. Studies evaluating KRAS mutation status in NSCLC patients have shown that mutations in the KRAS gene are strongly predictive of resistance to tyrosine kinase inhibitors such as Gifitinib and Erlotinib.

9. The Most Comprehensive 43 EGFR MUTATION PANEL available EGFR mutation testing along with other clinical factors, helps physicians guide treatment (Gifitinib/ Erlotinib) for patient with lung cancer. Samples Submission for EGFR Mutation Detection Tumor Samples •Routine practice and so-called “gold-standard” is to analyze DNA derived from a tumor sample, normally Formalin Fixed Paraffin Embedded (FFPET) diagnostic block. •DNA tends to be degraded, therefore, it is important to use robust well validated extraction methodologies to avoid assay fails and false negative results where possible.

10. Overview: CanAce-BR® assay that can help women with some types of breast cancer learn more about the biological activity of their specific tumor. Along with other pieces of information, the results from the assay can help women and their doctors make decisions about whether or not to include chemotherapy in their treatment plan. It can also help indicate how likely it is that a woman’s cancer may return in the future (distant reoccurrence). This assay looks at a group of 21 genes within a woman’s tumor sample—16 cancer genes and 5 control genes—to see how they are expressed, or how active they are. The results of the assay are reported as a quantitative Reoccurrence Score® result, which is a score between 0 and 100 that correlates with the likelihood of a woman’s chances of having her cancer return, and the likelihood that she will benefit from adding chemotherapy to her hormonal therapy. CanAce-BR® assay may in addition to standard measurements (such as tumor size, tumor grade and lymph node status) that doctors have traditionally used to estimate how likely a woman’s cancer is to return, and to help her make treatment decisions. Indications May be prescribed after surgery (lumpectomy or mastectomy), but before you and your doctor make a final decision regarding adjuvant treatment ASCO The Gene Based assay is recommended for use in newly diagnosed ER+, N- breast cancer patients to predict risk of recurrence. The assay can also be used to identify patients who may be successfully treated with tamoxifen and may not require adjuvant chemotherapy. “It has been suggested that tamoxifen-treated patients with an excellent estimated prognosis may be spared adjuvant chemotherapy.” Harris L, Fritsche H, Mennel R, et al. American Society of Clinical Oncology 2007 update of recommendations for the use of tumor markers in breast cancer. J ClinOncol. 2007;25(33):5287-312. NCCN “The option of using a gene-based assay of tumor tissue to help guide chemotherapy treatment decisions is now included within the systemic adjuvant treatment decision pathway for patients with node-negative or pN1mi (micrometastasis: 0.2 mm - 2.0 mm), hormone-receptor-positive, HER2-negative tumors that are 0.6 to 1.0 cm and moderately/poorly differentiated or with unfavorable features or > 1 cm.” METHODOLOGY AceProbe™ offers unparalleled detection rates using a Mass-Spectrometry Assay for Gene Analysis by multiplexed PCR on MALDI-TOF MS platform.

11. NO SIGNIFICANT BENEFIT WITH ADDITION OF CHEMO TO TAMOXIFEN SIGNIFICANT BENEFIT WITH ADDITION OF CHEMO TO TAMOXIFEN REFERENCES: Association Between the 21-Gene Recurrence Score Assay and Risk of Locoregional Recurrence in Node-Negative, Estrogen Receptor -Positive Breast Cancer: Results From NSABP B-14 and NSABP B-20. Eleftherios P. Mamounas,*Tang, Fisher, Paik, Shak, P. Costantino, Watson, E. Geyer Jr, Lawrence Wickerham, and Norman WolmarkJ ClinOncol. 2010; Prediction of Risk of Distant Recurrence Using the 21-Gene Recurrence Score in Node-Negative and Node-Positive Postmenopausal Breast Cancer Patients Treated with Anastrozole or Tamoxifen: A TransATAC Study Mitch Dowsett, Jack Cuzick, Christopher James Wale, John Forbes, Elizabeth A. Mallon, Janine Salter, Emma Quinn, Anita Dunbier, Mitch Baum, AmanBuzdar, Anthony Howell, Roberto Bugarini, Frederick L. Baehner, and Steven Shak J ClinOncol. 2010;

12. Gastrointestinal stromal tumor (GIST) is a rare cancer affecting the digestive tract or nearby structures within the abdomen. GI stromal tumor, or GIST cancer, is a sarcoma. Sarcomas are cancers that grow from cells of the body’s connective or supportive tissues such as bone, cartilage, tendons, nerves, fat, muscle, synovial tissue (tissue around joints), or blood vessels. (Most cancers are carcinomas, not sarcomas.) GIST cancers arise either from cells called Interstitial Cells of Cajal (ICCs) or from less differentiated stem cells or precursor cells that can develop into ICCs. The ICCs are known as “pacemaker cells of the gut” because they send signals to trigger peristalsis, the digestive tract’s muscular contractions that move food along its course. What causes GIST? Random genetic mutations are the apparent cause of GISTs. The majority of GISTs show identified mutations in cell-surface proteins called tyrosine kinase receptors. Most GISTs show mutations in a gene that produces a growth factor receptor called KIT. A few GISTs show mutations in the gene for a closely related receptor for platelet derived growth factor receptor alpha (PDGFR a or PDGFRA). . Where does GIST occur? Primary tumors are in the original site of development, before any spread to other locations. Primary gastrointestinal stromal tumors (GIST cancers) may occur anywhere along the gastrointestinal tract from the esophagus to the anus. The most frequent site for GISTs is the stomach (about 55%), followed by the duodenum and small intestine (about 30%), esophagus (about 5%), rectum (about 5%), colon (about 2%), and rare other locations. Occasionally, primary GISTs may develop in the supporting membranes of the abdominal organs (peritoneum, mesentery, omentum), the liver, the pancreas, the ovaries, the uterus, and the prostate. Because primary GISTs in these locations do not arise directly from the GI tract, they are sometimes called “extra gastrointestinal” stromal tumors (Reith et al, 2000). When a GIST grows in a location where it is not encased in the peritoneal membranes, it is said to be retroperitoneal. Cancer is most threatening when it metastasizes or spreads to additional locations beyond the primary tumor site. In GIST the most common sites for  metastasis are the liver and the abdominal membranes.  KIT mutations or PDGFRA mutations may cause GISTs

13. Treatments for GIST Surgery - Surgery is the first step in treatment for GIST, and it is often curative. Imatinib (Gleevec) - Imatinib (Gleevec) is FDA-approved for treatment of unresectable and metastatic GIST, as well as for prevention of recurrence of higher-risk GISTs (adjuvant therapy). Sunitinib (Sutent) - Sunitinib (Sutent) is FDA-approved for treatment of GIST resistant to Imatinib / Gleevec and for patients who are intolerant of Imatinib / Gleevec. Hepatic artery embolization - Embolization is a surgical procedure for treatment of liver metastases of GIST. Radiofrequency ablation - RFA is a surgical procedure for treatment of liver metastases of GIST. References Reddy P, Boci K, Charbonneau C. The epidemiologic, health-related quality of life, and economic burden of gastrointestinal stromal tumours. J ClinPharmTher. 2007;32:557-565. Miettinen M, Lasota J. Gastrointestinal stromal tumors. Review on morphology, molecular pathology, prognosis, and differential diagnosis. Arch Pathol Lab Med. 2006;130:1466-1478. Demetri GD, Benjamin RS, Blanke CD, et al. NCCN Task Force report: Management of patients with gastrointestinal stromal tumor (GIST)–Update of the NCCN clinical practice guidelines. J NatlComprCancNetw. 2007;5:s1-s28. Heinrich MC, Owzar K, Corless CL, et al. Correlation of kinase genotype and clinical outcome in the North American Intergroup Phase III Trial of imatinibmesylate for treatment of advanced gastrointestinal stromal tumor: CALGB 150105 study by Cancer and Leukemia Group B and Southwest Oncology Group. J ClinOncol. 2008;26:5360-5367. Debiec-Rychter M, Sciot R, Le Cesne A, et al. KIT mutations and dose selection for imatinib in patients with advanced gastrointestinal stromal tumours. Eur J Cancer. 2006;42:1093-1103. Blanke CD, Demetri GD, von Mehren M, et al. Long-term results from a randomized phase II trial of standard- versus higher-dose imatinibmesylate for patients with unresectable or metastatic gastrointestinal stromal tumors expressing KIT. J ClinOncol. 2008;26:620-625. Corless CL, Schroeder A, Griffith D, et al. PDGFRA mutations in gastrointestinal stromal tumors: frequency, spectrum and in vitro sensitivity to imatinib. J ClinOncol. 2005;23:5357-5364. Lasota J, Dansonka-Mieszkowska A, Sobin JH, et al. A great majority of GISTs with PDGFRA mutations represent gastric tumors of low or no malignant potential. Lab Invest. 2004;84:874-883. Demetri GD, van Oosterom AT, Garrett CR, et al. Efficacy and safety of sunitinib in patients with advanced gastrointestinal stromal tumour after failure of imatinib: a randomized controlled trial. Lancet. 2006;368:1329-1338. Specimen Requirements: Paraffin-embedded, formalin-fixed tissue block, Protect paraffin block from excessive heat. Ship in cooled container during summer.Remarks: Only tissue that is clearly tumor (established by histological criteria) should be tested.

14. Tamoxifen resistance DNA test – CYP2D6 Tamoxifen, due to its low-cost and low-side-effect profile, is the drug of choice for patients with ER+ breast cancer. Reduces recurrence by 50% and mortality by one-third at 15-year follow up • CYP2D6 genotype associations for metabolism • Poor metabolizer (PM) • Two nonfunctional CYP2D6 alleles • Highly impaired metabolism of the drug, causing lack of drug response • Intermediate metabolizer (IM) • One nonfunctional CYP2D6 allele and either one functional or decreased-functional allele, or 2 decreased-functional alleles • Possible impaired metabolism of the drug • Extensive metabolizer (EM) • 2 functional CYP2D6 alleles, or one copy each of a functional and a decreased-functional allele • Normal metabolism • Ultrarapid metabolizer (UM) • >2 functional CYP2D6 alleles • Increased metabolism; consider not using drugs metabolized by the CYP2D6 gene • Approximately 5% of Caucasians are predicted to have UM phenotypes Tamoxifen is an anti-estrogen drug used in the treatment of estrogen-receptor positive (ER+) breast cancer to reduce the risk of recurrence. Mutations in CYP2D6 may lead to altered drug metabolism and reduce the concentration of active metabolite (e.g, endoxifen) available. The reduction in active metabolite concentrations may reduce likelihood of response, manifested by an increased risk of recurrence of breast cancer. Mutation detection include: Sample submission: * Collect 5 ml blood from adult in an EDTA anticoagulant tube * Store-refrigerate blood sample * Ship under 2-8 degree * Receipt by the laboratory within 24 hours of collection CYP2D6*4 (1846G>A) and CYP2D6*10 (100C>T)

15. JAK2 Mutation Analysis in Myeloproliferative Disorders Investigating for point mutation V617F Clinical Indications Distinguishing MPD from other conditions (such as secondary thrombocytosis ) can be difficult and has been based primarily on the clinical criteria. These criteria are ambiguous and at times misleading. Identification of the single point mutations in JAK2 allows for reliable diagnosis, classification and potential treatment for MPDs.

16. KRAS Role in Colorectal Cancer • KRAS is a gene that encodes one of the proteins in EGFR signaling pathway. • KRAS protein plays a central role in tumor development, regulating down stream proteins that are involved in proliferation, survival, metastasis and angiogenesis. • The KRAS status of a tumor may be indicative of prognostic and predictive response to certain drugs. • Clinical significance of KRAS mutations • The presence of somatic mutations in the KRAS gene is associated with poor prognosis and non-response to anti-EGFR therapy in colorectal and non-small cell lung cancer patients. • Studies evaluating KRAS mutation status in colorectal patients demonstrate that KRAS mutations are strongly correlated with lack of response to cetuximab and panitumumab, shorter progression-free survival (PFS) and shorter overall survival. • Studies evaluating KRAS mutation status in NSCLC patients have shown that mutations in the KRAS gene are strongly predictive of resistance to tyrosine kinase inhibitors such as Gifitinib and Erlotinib. Testing of KRAS status allows a patient to receive treatment that is personalized to them.

17. AceProbe™ offers a comprehensive KRAS mutation test menu to assist with personalized test management of patients with metastatic colorectal cancer. AceProbe™ offers unparalleled detection rates using a Mass-Spectrometry assay for codons 12 and 13 of KRAS gene by SNP extension n MALDI-TOF MS platform. Five additional codon 13 mutations increases coverage REFERENCES: 1. Lievre A., et al. Cancer Res (2006), 66: 3992-3995;2. Di Fiore F., et al. Br J Cancer (2007), 96:1166-1169; 3. Lièvre A., et al. JCO May (2008), 20: 2601-2602; 4. Lievre A., et al. JCO (2008), 26: 374-379; 5. Amado R.G., et al. J Clin Oncol (2008), 26:1626-1634; 6. De Roock W., et al. Ann Oncol (2008), 19:508-515; 7. Khambata-Ford S., et al. J Clin Oncol (2008), 25:3230-3237; 8. Freeman D.J., et al. Clin Colorectal Cancer (2008), 7:184-190; 9. Di Nicolantonio F. et al. J Clin Oncol (2008). 26(35):5705-12;10. Loupakis F., et al. Br J Cancer (2009). 101(4):715-21; 11. Van Custem E. ASCO GI 2010. Abstract 281; 12. Benvenuti, S., et al. Cancer Res(2007). 67(6): p. 2643-48; 13. Font, A., et al. Dis Colon Rectum (2001). 44(4): p. 549-57; 14. Freeman, D.J., et al. Clin Colorectal Cancer (2008). 7(3): p. 184-90 15. Karapetis, C.S., et al. N Engl J Med (2008). 359(17): p. 1757-65 16. Lee, J.C., et al. AnticancerRes (1996). 16(6B): p. 3839-44 17. Tanaka, M., et al. J SurgOncol (1994). 57(1): p. 57-64 18. Zhu, C.Q., et al. J Clin Oncol (2008). 26(26): p. 4268-75 19. Scheffzek, K.,et al. Trends BiochemSci,(1998). 23(7): p. 257-62 • Specimen requirement • Formalin Fixed Paraffin Embedded (FFPET) diagnostic block. • Store specimen at room temperature (25 degree C)

18. The great individual variability in both outcome and toxicity of platinum chemotherapy requires the identification of genetic markers. Platinum chemo agents have been the treatment of choice for ovarian cancer for the past 20 years. Now they are also proving effective against certain other cancers including testicular, bladder, endometrial, colon, and lung cancer and some cancers of the head and neck. Platinum chemotherapy drugs are given as an intravenous drip. In many cases, Cisplatin and Carboplatin are given in combination with other chemotherapy drugs. For example, most women with ovarian cancer are treated with either Cisplatin or Carboplatin alone, or one of these in combination with paclitaxel (Taxol). Oxaliplatin is commonly used to treat metastatic colon or rectal cancer, often in combination with fluorouracil or leukovorin. Chemo agents containing platinum can be very effective in fighting aggressive cancers such as ovarian cancer and late-stage lung and colon cancer, but they also cause many unpleasant side effects such as nausea, fatigue, dry mouth, and neuropathy. They can also cause infertility and kidney damage. In addition, studies have shown that patients who take Cisplatin and Carboplatin have an elevated risk of developing leukemia down the line. Sample submission: * Collect 5 ml blood from adult in an EDTA anticoagulant tube * Store-refrigerate blood sample * Ship under 2-8 degree * Receipt by the laboratory within 24 hours of collection METHODOLOGY AceProbe® offers unparalleled detection rates using a Mass-Spectrometry Assay for Gene Analysis by multiplexed PCR on MALDI-TOF MS platform.

19. Indications: To investigate polymorphisms with putative influence on Platinum Chemo Agent activity and their association with clinical outcomes of patients with cancers. References: Lecomte T, Landi B, Beaune P et al. Glutathione S-transferase P1 polymorphism (Ile105Val) predicts cumulative neuropathy in patients receiving oxaliplatin-based chemotherapy. Clin Cancer Res 2006; 12: 3050–3056.

20. Sensitive detection and quantification of High- Risk Human Papillomavirus types . • References: • 1.) Lo et al., Cancer Res. 2000 60 (24): 6878‐6881. • 2.) Capone et al., Clin. Cancer Res. 2000 (6): 4171‐4175. • 3.) Ha et al., Clin. Cancer Res. 2002 (8): 1203‐1209. • 4.) Yang et al., PNAS 2005 (21): 102, 7683‐7688. • 5.) Choi YD, Jung WW, Nam JH, Choi HS, Park CS. Detection of HPV genotypes in cervical lesions by the HPV DNA chip and sequencing. GynecolOncol 2005;98: 369-375. • 6.) Patel DA, Shih Y, Newton DW, Michael CW, Oeth PA, Kane MD, Opipari AW, Ruffin MT, Kalikin LM, 7.) Kurnit DM. Development and Evaluation of a PCR and Mass Spectrometry-based (PCR-MS) Method for Quantitative, Type-specific Detection of Human Papillomavirus, J Virol Methods 2009;160:78-84. • 8.) Yang H, Yang K, Khafagi A, Tang Y, Carey TE, Opipari AW, Lieberman R, Oeth PA, Lancaster W, Klinger HP, Kaseb AO, Metwally A, Khaled H, Kurnit DM. Sensitive detection of human papillomavirus in cervical, head/neck, and schistosomiasis-associated bladder malignancies. Proc NatlAcadSci 2005;102:7683-8. • 9.) Munagala R, Gabriella Dona M, Rai SN, Jenson AB, Bala N, Ghim SJ, Gupta RC. Sihgnificance of multiple HPV infection in cervical cancer patients and its impact on treatment response. Int J Oncol 2009;34:263-71. METHODOLOGY AceProbe™ offers unparalleled detection rates using a Mass-Spectrometry assay for HPV DETECTION • All 15 oncogenic HPV types • Genotype specific results • Any Sample Source- Blood Tissue- serum • Ultra high Sensitivity and specificity • Handles low volume samples • Fully Automated high throughput • Option of Quantitative viral Load Estimation.

21. Beta – Thalassemia Confirmatory Test- 14 Mutations Test With coverage of 95% mutations Beta thalassemiais due to mutations in one or both of the beta globin genes. There are 100 to 200 mutations that have been identified but only about 14 are commonly reported for Indian Populations. The severity of the anemia caused by beta thalassemia depends on which mutations are present and on whether they decrease beta globin production (called beta+ thalassemia) or completely eliminate it (called beta0 thalassemia). The different types of beta thalassemia include: Beta Thalassemia Trait. A person with this condition has one normal gene and one with a mutation. They will usually experience no health problems other than microcytosis and a possible mild anemia that will not respond to iron supplements. This gene mutation can be passed on to an individual’s children. Thalassemia Intermedia. In this condition, an affected person has two abnormal genes but is still producing some beta globin. The severity of the anemia and health problems experienced depends on the mutations present. The dividing line between thalassemia intermedia and thalassemia major is the degree of anemia and the number and frequency of blood transfusions required to treat it. Those with thalassemia intermedia may need occasional transfusions but do not require them on a regular basis. Thalassemia Major (also called Cooley's Anemia). This is the most severe form of beta thalassemia. The patient has two abnormal genes that cause either a severe decrease or complete lack of beta globin production, preventing the production of significant amounts of Hb A. This condition usually appears in an infant after three months of age and causes life-threatening anemia. This anemia requires lifelong regular blood transfusions and considerable ongoing medical care. Over time, these frequent transfusions lead to excessive amounts of iron in the body. Left untreated, this excess iron can deposit into the liver, heart and other organs and can lead to a premature death from organ failure. Inheritance: Thalassemia major and thalassemia intermedia are inherited in an autosomal recessive pattern, which means both copies of the HBB gene in each cell have mutations. The parents of an individual with an autosomal recessive condition each carry one copy of the mutated gene, but they typically do not show signs and symptoms of the condition. Sometimes, however, people with only one HBB gene mutation in each cell develop mild anemia. These mildly affected people are said to have thalassemia minor. In a small percentage of families, the HBB gene mutation is inherited in an autosomal dominant manner. In these cases, one copy of the altered gene in each cell is sufficient to cause the signs and symptoms of beta thalassemia.

22. BENEFIT : DNA analysis. This test is used to investigate deletions and mutations in the alpha and beta globin producing genes. Family studies have been done to evaluate carrier status and the types of mutations present in other family members. DNA testing is routinely done and can be used to help diagnose thalassemia and to determine carrier status. AceProbe mutation panel provides a complete diagnosis by analyzing almost all mutations to include a ~ 619bp deletion in the same assay. Sample submission: * Collect 3ml from child or 5 ml blood from adult in an EDTA anticoagulant tube * Store-refrigerate blood sample * Ship under 2-8 degree * Receipt by the laboratory within 24 hours of collection

24. The AceProbe® Difference AceProbe® Genetic Diagnostics provides physicians with genetics based testing and prognosis solutions that identify a patient’s predisposition to major diseases and the therapeutic considerations that can improve treatment outcomes and quality of life. Our initial solutions have focused on enhancing the safety and effectiveness of anticoagulation drugs, including two of the world’s most widely prescribed medications for patients at risk of stroke and other serious circulatory problems. These timely and cost effective tests can help to significantly reduce adverse bleeding and other events in patients that could benefit from dosage adjustments or alternative therapies. At AceProbe® we offer: • Fast turnaround: Samples are quickly processed and results reported, normally within committed time. • Understandable reports: Test results are clearly presented with the needs of the medical professional in mind. • Comprehensive clinical support: Should you have any questions or need more detailed information regarding test results, there are PhD level support personnel waiting for your call. For more information & Sample Collection Contact: AceProbe Technologies (India) Pvt. Ltd., B-8, Namdhari Chambers, 9/54, D.B. Gupta Road, Karol Bagh, New Delhi-110005 Phone: +91-11-45098118 Fax: +91-11-23552378 Email: info@aceprobe.com; Website: www.aceprobe.com