Skip to main content Accessibility help

We use cookies to distinguish you from other users and to provide you with a better experience on our websites. Close this message to accept cookies or find out how to manage your cookie settings.

Close cookie message
×
Hostname: page-component-669899f699-swprf Total loading time: 0 Render date: 2025-04-24T14:31:21.204Z Has data issue: false hasContentIssue false

7 - Miniaturized polymerase chain reaction for quantitative clinical diagnostics

Published online by Cambridge University Press:  25 January 2011

By
Melissa Mariani ,
Lin Chen  and
Philip J. Day
Edited by
Stephen A. Bustin
Stephen A. Bustin
Affiliation:
Queen Mary University of London
Get access

Summary

CURRENT QUANTITATIVE POLYMERASE CHAIN REACTION USES IN QUANTITATIVE CLINICAL DIAGNOSTICS

Clinical diagnostics is progressively embracing the incorporation of translational medicine, as the diagnostic industry starts to lean toward personalized treatment. Direct benefits to clinical diagnostics will be achieved as a result of technical advancements providing unambiguous quantitative analysis of the transcriptome, although the quantitative clinical diagnostic sector is a relatively small part of the overall clinical diagnostic market. Interests in deoxyribonucleic acid (DNA) and messenger ribonucleic acid (mRNA) detection and quantification have improved current knowledge of cell functions, including cell regulation, growth, expression markers, and transcription. The polymerase chain reaction (PCR) is one such research technique in mainstream clinical diagnostics that can provide quantitative analysis and assist in closing the “bench to bedside” gap found with translational medicine.

The reverse transcription–PCR (RT–PCR) is at present the most sensitive technique for mRNA detection, although quantitative RT–PCR (qRT–PCR) increasingly provides robust PCR product measurements from each cycle. Additionally, qPCR is commonly applied to clinical diagnostics, making the technique an industry standard for RNA product detection and quantification. An increase in qPCR applications, combined with the growing importance of clinical diagnostics, has permitted both areas to develop in parallel. Reflecting these advancements, the value of qPCR in quantitative clinical diagnostics has increased largely through the stochastic integration of fluorochrome that can be directly related to qualitative measurements. Qualitative end-point PCR measurements benefit from many of the detection strategies used in qPCR, but obviously lack quantitative application.

Type
Chapter
Information
The PCR Revolution
Basic Technologies and Applications
 , pp. 88 - 109
Publisher: Cambridge University Press
Print publication year: 2009

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

Book purchase

Temporarily unavailable

References

Bustin, SA (2002) Quantification of mRNA using real-time reverse transcription PCR (RT-PCR): trends and problems. Journal of Molecular Endocrinology 29(1): 23–39.CrossRefGoogle ScholarPubMed
Ramachandran, C, Melnick, SJ (1999) Multidrug resistance in human tumors – molecular diagnosis and clinical significance. Journal of Molecular Diagnostics 4(2): 81–94.Google ScholarPubMed
Burrow, GN (1988) From bench to bedside – the impact of the transfer of the new biology to clinical medicine. Clinical and Investigative Medicine 11(4): 315–320.Google ScholarPubMed
Bustin, SA (2000) Absolute quantification of mRNA using real-time reverse transcription polymerase chain reaction assays. Journal of Molecular Endocrinology 25(2): 169–193.CrossRefGoogle ScholarPubMed
Bustin, SA, Nolan, T (2004) Pitfalls of quantitative real-time reverse-transcription polymerase chain reaction. Journal of Biomolecular Techniques 15(3): 155–166.Google ScholarPubMed
Ginzinger, DG (2002) Gene quantification using real-time quantitative PCR: an emerging technology hits the mainstream. Experimental Hematology 30(6): 503–512.CrossRefGoogle Scholar
Halford, WP, Falco, VC, et al. (1999) The inherent quantitative capacity of the reverse transcription-polymerase chain reaction. Analytical Biochemistry 266(2): 181–191.CrossRefGoogle ScholarPubMed
Bustin, SA, Benes, V, et al. (2005) Quantitative real-time RT-PCR – a perspective. Journal of Molecular Endocrinology 34(3): 597–601.CrossRefGoogle ScholarPubMed
Mackay, IM, Arden, KE, et al. (2002) Real-time PCR in virology. Nucleic Acids Research 30(6): 1292–1305.CrossRefGoogle ScholarPubMed
Mackay, J, Landt, O (2007) Real-time PCR fluorescent chemistries. Methods in Molecular Biology (Clifton, N.J.) 353: 237–261.Google ScholarPubMed
Dong, QH, Zheng, S, et al. (2005) Evaluation of ST13 gene expression in colorectal cancer patients. Journal of Zhejiang University. Science B 6(12): 1170–1175.CrossRefGoogle ScholarPubMed
Hill, WE (1996) The polymerase chain reaction: applications for the detection of foodborne pathogens. Critical Reviews in Food Science and Nutrition 36(1–2): 123–173.CrossRefGoogle ScholarPubMed
Hoebeeck, J, Speleman, F, et al. (2007) Real-time quantitative PCR as an alternative to Southern blot or fluorescence in situ hybridization for detection of gene copy number changes. Methods in Molecular Biology (Clifton, N.J.) 353: 205–226.Google ScholarPubMed
Jolkowska, J, Derwich, K, et al. (2007) Methods of minimal residual disease (MRD) detection in childhood haematological malignancies. Journal of Applied Genetics 48(1): 77–83.CrossRefGoogle ScholarPubMed
Ratcliff, RM, Chang, G, et al. (2007) Molecular diagnosis of medical viruses. Current Issues in Molecular Biology 9(2): 87–102.Google ScholarPubMed
Stahlberg, A, Hakansson, J, et al. (2004) Properties of the reverse transcription reaction in mRNA quantification. Clinical Chemistry 50(3): 509–515.CrossRefGoogle ScholarPubMed
Sims, CE, Allbritton, NL (2007) Analysis of single mammalian cells on-chip. Lab on a Chip 7(4): 423–440.CrossRefGoogle ScholarPubMed
Schutten, M, Niesters, HG (2001) Clinical utility of viral quantification as a tool for disease monitoring. Expert Review of Molecular Diagnostics 1(2): 153–162.CrossRefGoogle ScholarPubMed
Kao, CL, King, CC, et al. (2005) Laboratory diagnosis of dengue virus infection: current and future perspectives in clinical diagnosis and public health. Journal of Microbiology, Immunology, and Infection 38(1): 5–16.Google ScholarPubMed
Templeton, KE, Scheltinga, SA, et al. (2004) Rapid and sensitive method using multiplex real-time PCR for diagnosis of infections by influenza A and influenza B viruses, respiratory syncytial virus, and parainfluenza viruses 1, 2, 3, and 4. Journal of Clinical Microbiology 42(4): 1564–1569.CrossRefGoogle Scholar
Mulder, J, McKinney, N, et al. (1994) Rapid and simple PCR assay for quantitation of human immunodeficiency virus type 1 RNA in plasma: application to acute retroviral infection. Journal of Clinical Microbiology 32(2): 292–300.Google ScholarPubMed
Sun, R, Ku, J, et al. (1998) Ultrasensitive reverse transcription-PCR assay for quantitation of human immunodeficiency virus type 1 RNA in plasma. Journal of Clinical Microbiology 36(10): 2964–2969.Google ScholarPubMed
Palmer, S, Wiegand, AP, et al. (2003) New real-time reverse transcriptase-initiated PCR assay with single-copy sensitivity for human immunodeficiency virus type 1 RNA in plasma. Journal of Clinical Microbiology 41(10): 4531–4536.CrossRefGoogle ScholarPubMed
Candotti, D, Temple, J, et al. (2004) Multiplex real-time quantitative RT-PCR assay for hepatitis B virus, hepatitis C virus, and human immunodeficiency virus type 1. Journal of Virological Methods 118(1): 39–47.CrossRefGoogle ScholarPubMed
Eder, M, Battmer, K, et al. (1999) Monitoring of BCR-ABL expression using real-time RT-PCR in CML after bone marrow or peripheral blood stem cell transplantation. Leukemia 13(9): 1383–1389.CrossRefGoogle ScholarPubMed
Emig, M, Saussele, S, et al. (1999) Accurate and rapid analysis of residual disease in patients with CML using specific fluorescent hybridization probes for real time quantitative RT-PCR. Leukemia 13(11): 1825–1832.CrossRefGoogle ScholarPubMed
Gabert, J (1999) Detection of recurrent translocations using real time PCR; assessment of the technique for diagnosis and detection of minimal residual disease. Haematologica 84 Suppl EHA-4: 107–109.Google ScholarPubMed
Hochhaus, A, Reiter, A, et al. (1998) Molecular monitoring of residual disease in chronic myelogenous leukemia patients after therapy. Recent Results in Cancer Research 144: 36–45.CrossRefGoogle ScholarPubMed
Olavarria, E, Kanfer, E, et al. (2001) Early detection of BCR-ABL transcripts by quantitative reverse transcriptase-polymerase chain reaction predicts outcome after allogeneic stem cell transplantation for chronic myeloid leukemia. Blood 97(6): 1560–1565.CrossRefGoogle ScholarPubMed
Szczepanski, T. (2007) Why and how to quantify minimal residual disease in acute lymphoblastic leukemia?Leukemia 21(4): 622–626.CrossRefGoogle ScholarPubMed
Tysarowski, A, Fabisiewicz, A, et al. (2007) Usefulness of real-time PCR in long-term follow-up of follicular lymphoma patients. Acta Biochimica Polonica 54(1): 135–142.Google ScholarPubMed
Park, NJ, Zhou, X, et al. (2007) Characterization of salivary RNA by cDNA library analysis. Archives of Oral Biology 52(1): 30–35.CrossRefGoogle ScholarPubMed
Muller, MC, Hordt, T, et al. (2004) Standardization of preanalytical factors for minimal residual disease analysis in chronic myelogenous leukemia. Acta Haematologica 112(1–2): 30–33.CrossRefGoogle ScholarPubMed
Raengsakulrach, B, Nisalak, A, et al. (2002) Comparison of four reverse transcription-polymerase chain reaction procedures for the detection of dengue virus in clinical specimens. Journal of Virological Methods 105(2): 219–232.CrossRefGoogle ScholarPubMed
Keilholz, U, Willhauck, M, et al. (1998) Reliability of reverse transcription-polymerase chain reaction (RT-PCR)-based assays for the detection of circulating tumour cells: a quality-assurance initiative of the EORTC Melanoma Cooperative Group. European Journal of Cancer 34(5): 750–753.CrossRefGoogle ScholarPubMed
Nolan, T, Hands, RE, et al. (2006) Quantification of mRNA using real-time RT-PCR. Nature Protocols 1(3): 1559–1582.CrossRefGoogle ScholarPubMed
Koh, CG, Tan, W, et al. (2003) Integrating polymerase chain reaction, valving, and electrophoresis in a plastic device for bacterial detection. Analytical Chemistry 75(17): 4591–4598.CrossRefGoogle Scholar
Afzal, M (2003) Comparative evaluation of measles virus-specific RT-PCR methods through an international collaborative study. Journal of Medical Virology 70: 171–176.CrossRefGoogle ScholarPubMed
Niesters, HG (2001) Quantitation of viral load using real-time amplification techniques. Methods 25(4): 419–429.CrossRefGoogle ScholarPubMed
Niesters, HG (2004) Molecular and diagnostic clinical virology in real time. Clinical Microbiology and Infection 10(1): 5–11.CrossRefGoogle Scholar
Kwok, S, Higuchi, R (1989) Avoiding false positives with PCR. Nature 339(6221): 237–238.CrossRefGoogle ScholarPubMed
Bustin, SA, Dorudi, S (2004) Gene expression profiling for molecular staging and prognosis prediction in colorectal cancer. Expert Review of Molecular Diagnostics 4(5): 599–607.CrossRefGoogle ScholarPubMed
Bustin, SA, Siddiqi, S, et al. (2004) Quantification of cytokeratin 20, carcinoembryonic antigen and guanylyl cyclase C mRNA levels in lymph nodes may not predict treatment failure in colorectal cancer patients. International Journal of Cancer 108(3): 412–417.CrossRefGoogle Scholar
Schuster, R, Max, N, et al. (2004) Quantitative real-time RT-PCR for detection of disseminated tumor cells in peripheral blood of patients with colorectal cancer using different mRNA markers. International Journal of Cancer 108(2): 219–227.CrossRefGoogle ScholarPubMed
Akane, A, Matsubara, K, et al. (1994) Identification of the heme compound copurified with deoxyribonucleic acid (DNA) from bloodstains, a major inhibitor of polymerase chain reaction (PCR) amplification. Journal of Forensic Science 39(2): 362–372.CrossRefGoogle Scholar
Bustin, SA (2005) Real-time, fluorescence-based quantitative PCR: a snapshot of current procedures and preferences. Expert Review of Molecular Diagnostics 5(4): 493–498.CrossRefGoogle ScholarPubMed
Wolffs, P, Norling, B, et al. (2005) Risk assessment of false-positive quantitative real-time PCR results in food, due to detection of DNA originating from dead cells. Journal of Microbiological Methods 60(3): 315–323.CrossRefGoogle ScholarPubMed
Bomjen, G, Raina, A, et al. (1996) Effect of storage of blood samples on DNA yield, quality and fingerprinting: a forensic approach. Indian Journal of Experimental Biology 34(4): 384–386.Google ScholarPubMed
Bertolini, E, Olmos, A, et al. (2001) Single-step multiplex RT-PCR for simultaneous and colourimetric detection of six RNA viruses in olive trees. Journal of Virological Methods 96(1): 33–41.CrossRefGoogle ScholarPubMed
Curry, J, McHale, C, Smith, MT (2002) Low efficiency of the Moloney murine leukemia virus reverse transcriptase during reverse transcription of rare t(8;21) fusion gene transcripts. BioTechniques 32(4): 768, 770, 772, 754–755.Google Scholar
Day, PJ (2006) Miniaturization applied to analysis of nucleic acids in heterogeneous tissues. Expert Review of Molecular Diagnostics 6(1): 23–28.CrossRefGoogle ScholarPubMed
Wittwer, CT, Herrmann, MG, et al. (1997) Continuous fluorescence monitoring of rapid cycle DNA amplification. BioTechniques 22(1): 130–131, 134–138.Google ScholarPubMed
Yin, JS, Shackel, NA, Zekry, A, McGuinness, PH, Richards, C, Putten, KV, et al. (2001) Real-time reverse-transcriptase polymerase chain reaction (RT-PCR) for measurement of cytokine and growth factor mRNA expression with fluorogenic probes or SYBR Green I. Immunology and Cell Biology 79: 213.CrossRefGoogle ScholarPubMed
Ogura, M, Mitsuhashi, M (1994) Screening method for a large quantity of polymerase chain reaction products by measuring YOYO-1 fluorescence on 96-well polypropylene plates. Analytical Biochemistry 218(2): 458–459.CrossRefGoogle ScholarPubMed
Hoerndli, FJ, Toigo, M, et al. (2004) Reference genes identified in SH-SY5Y cells using custom-made gene arrays with validation by quantitative polymerase chain reaction. Analytical Biochemistry 335(1): 30–41.CrossRefGoogle ScholarPubMed
Greiner, O, Day, PJ, et al. (2001) Quantitative detection of Streptococcus pneumoniae in nasopharyngeal secretions by real-time PCR. Journal of Clinical Microbiology 39(9): 3129–3134.CrossRefGoogle ScholarPubMed
Greiner, O, Day, PJ, et al. (2003) Quantitative detection of Moraxella catarrhalis in nasopharyngeal secretions by real-time PCR. Journal of Clinical Microbiology 41(4): 1386–1390.CrossRefGoogle ScholarPubMed
Schulte, TH, Bardell, RL, et al. (2002) Microfluidic technologies in clinical diagnostics. Clinica Chimica Acta 321(1–2): 1–10.CrossRefGoogle ScholarPubMed
Sia, SK, Linder, V, et al. (2004) An integrated approach to a portable and low-cost immunoassay for resource-poor settings. Angewandte Chemie (International ed. in English) 43(4): 498–502.CrossRefGoogle ScholarPubMed
Thorsen, T, Maerkl, SJ, et al. (2002) Microfluidic large-scale integration. Science 298(5593): 580–584.CrossRefGoogle ScholarPubMed
Steigert, J, Grumann, M, et al. (2006) Fully integrated whole blood testing by real-time absorption measurement on a centrifugal platform. Lab on a Chip 6(8): 1040–1044.CrossRefGoogle ScholarPubMed
Steigert, J, Grumann, M, Dube, M, Streule, W, Riegger, L, Brenner, T, et al. (2006) Direct hemoglobin measurement on a centrifugal microfluidic platform for point-of-care diagnostics. Sensors and Actuators A–Physical 130–131: 228–233.CrossRefGoogle Scholar
Auroux, PA, Iossifidis, D, et al. (2002) Micro total analysis systems. 2. Analytical standard operations and applications. Analytical Chemistry 74(12): 2637–2652.CrossRefGoogle ScholarPubMed
Auroux, PA, Koc, Y, et al. (2004) Miniaturised nucleic acid analysis. Lab on a Chip 4(6): 534–546.CrossRefGoogle ScholarPubMed
Dittrich, PS, Tachikawa, K, et al. (2006) Micro total analysis systems. Latest advancements and trends. Analytical Chemistry 78(12): 3887–3908.CrossRefGoogle ScholarPubMed
Kricka, LJ, Wilding, P (2003) Microchip PCR. Analytical and Bioanalytical Chemistry 377(5): 820–825.CrossRefGoogle ScholarPubMed
Roper, MG, Easley, CJ, et al. (2005) Advances in polymerase chain reaction on microfluidic chips. Analytical Chemistry 77(12): 3887–3893.CrossRefGoogle ScholarPubMed
Vilkner, T, Janasek, D, et al. (2004) Micro total analysis systems. Recent developments. Analytical Chemistry 76(12): 3373–3385.CrossRefGoogle ScholarPubMed
Reyes, DR, Iossifidis, D, et al. (2002) Micro total analysis systems. 1. Introduction, theory, and technology. Analytical Chemistry 74(12): 2623–2636.CrossRefGoogle Scholar
Zhang, CS, Xu, JL, et al. (2006) PCR microfluidic devices for DNA amplification. Biotechnology Advances 24(3): 243–284.CrossRefGoogle ScholarPubMed
Wittwer, CT, Garling, DJ (1991) Rapid cycle DNA amplification: time and temperature optimization. BioTechniques 10(1): 76–83.Google ScholarPubMed
Burns, MA, Mastrangelo, CH, et al. (1996) Microfabricated structures for integrated DNA analysis. Proceedings of the National Academy of Sciences of the United States of America 93(11): 5556–5561.CrossRefGoogle ScholarPubMed
Cheng, J, Shoffner, MA, et al. (1996) Chip PCR. 2. Investigation of different PCR amplification systems in microfabricated silicon-glass chips. Nucleic Acids Research 24(2): 380–385.CrossRefGoogle Scholar
Cheng, J, Waters, LC, et al. (1998) Degenerate oligonucleotide primed polymerase chain reaction and capillary electrophoretic analysis of human DNA on microchip-based devices. Analytical Biochemistry 257(2): 101–106.CrossRefGoogle ScholarPubMed
Kopp, MU, Mello, AJ, et al. (1998) Chemical amplification: continuous-flow PCR on a chip. Science 280(5366): 1046–1048.CrossRefGoogle ScholarPubMed
Waters, LC, Jacobson, SC, et al. (1998) Microchip device for cell lysis, multiplex PCR amplification, and electrophoretic sizing. Analytical Chemistry 70(1): 158–162.CrossRefGoogle ScholarPubMed
Waters, LC, Jacobson, SC, et al. (1998) Multiple sample PCR amplification and electrophoretic analysis on a microchip. Analytical Chemistry 70(24): 5172–5176.CrossRefGoogle ScholarPubMed
Ueda, M, Nakanishi, H, et al. (2000) Imaging of a band for DNA fragment migrating in microchannel on integrated microchip. Materials Science & Engineering C–Biomimetic and Supramolecular Systems 12(1–2): 33–36.CrossRefGoogle Scholar
Chen, L, Ren, J (2004) High-throughput DNA analysis by microchip electrophoresis. Combinatorial Chemistry & High Throughput Screening 7(1): 29–43.CrossRefGoogle ScholarPubMed
Obeid, PJ, Christopoulos, TK (2003) Continuous-flow DNA and RNA amplification chip combined with laser-induced fluorescence detection. Analytica Chimica Acta 494(1–2): 1–9.CrossRefGoogle Scholar
Ibrahim, MS, Lofts, RS, et al. (1998) Real-time microchip PCR for detecting single-base differences in viral and human DNA. Analytical Chemistry 70(9): 2013–2017.CrossRefGoogle ScholarPubMed
Northrup, MA, Benett, B, et al. (1998) A miniature analytical instrument for nucleic acids based on micromachined silicon reaction chambers. Analytical Chemistry 70(5): 918–922.CrossRefGoogle ScholarPubMed
Cady, NC, Stelick, S, et al. (2005) Real-time PCR detection of Listeria monocytogenes using an integrated microfluidics platform. Sensors and Actuators B–Chemical 107(1): 332–341.CrossRefGoogle Scholar
Xiang, Q, Xu, B, et al. (2005) Real time PCR on disposable PDMS chip with a miniaturized thermal cycler. Biomedical Microdevices 7(4): 273–279.CrossRefGoogle ScholarPubMed
Hu, GQ, Xiang, Q, et al. (2006) Electrokinetically controlled real-time polymerase chain reaction in microchannel using Joule heating effect. Analytica Chimica Acta 557(1–2): 146–151.CrossRefGoogle Scholar
Nakayama, T, Kurosawa, Y, et al. (2006) Circumventing air bubbles in microfluidic systems and quantitative continuous-flow PCR applications. Analytical and Bioanalytical Chemistry 386(5): 1327–1333.CrossRefGoogle ScholarPubMed
Zhou, ZM, Liu, DY, et al. (2004) Determination of SARS-coronavirus by a microfluidic chip system. Electrophoresis 25(17): 3032–3039.CrossRefGoogle ScholarPubMed
Kaigala, GV, Huskins, RJ, et al. (2006) Automated screening using microfluidic chip-based PCR and product detection to assess risk of BK virus-associated nephropathy in renal transplant recipients. Electrophoresis 27(19): 3753–3763.CrossRefGoogle ScholarPubMed
Liu, CN, Toriello, NM, et al. (2006) Multichannel PCR-CE microdevice for genetic analysis. Analytical Chemistry 78(15): 5474–5479.CrossRefGoogle ScholarPubMed
Lee, TM, Hsing, IM, Lao, AI, Carles, MC (2000) A miniaturized DNA amplifier: Its application in traditional Chinese medicine. Analytical Chemistry 72(17): 4242–4247.CrossRefGoogle ScholarPubMed
Giordano, BC, Copeland, ER, et al. (2001) Towards dynamic coating of glass microchip chambers for amplifying DNA via the polymerase chain reaction. Electrophoresis 22(2): 334–340.3.0.CO;2-O>CrossRefGoogle ScholarPubMed
Lee, TM, Carles, MC, et al. (2003) Microfabricated PCR-electrochemical device for simultaneous DNA amplification and detection. Lab on a Chip 3(2): 100–105.CrossRefGoogle Scholar
Liu, RH, Yang, JN, et al. (2004) Self-contained, fully integrated biochip for sample preparation, polymerase chain reaction amplification, and DNA microarray detection. Analytical Chemistry 76(7): 1824–1831.CrossRefGoogle ScholarPubMed
Lotspeich, E, Schoene, M, et al. (2007) Detection of disseminated tumor cells in the lymph nodes of colorectal cancer patients using a real-time polymerase chain reaction assay. Langenbeck's Archives of Surgery 392(5): 559–566.CrossRefGoogle ScholarPubMed
Snow, M, McKay, P, et al. (2006) Development, application and validation of a Taqman real-time RT-PCR assay for the detection of infectious salmon anaemia virus (ISAV) in Atlantic salmon (Salmo salar). Developments in Biologicals 126: 133–145; discussion 325–326.Google Scholar
Oberg, AN, Lindmark, GE, et al. (2004) Detection of occult tumour cells in lymph nodes of colorectal cancer patients using real-time quantitative RT-PCR for CEA and CK20 mRNAS. International Journal of Cancer 111(1): 101–110.CrossRefGoogle ScholarPubMed
Tsimberidou, AM, Jiang, Y, et al. (2002) Quantitative real-time polymerase chain reaction for detection of circulating cells with t(14;18) in volunteer blood donors and patients with follicular lymphoma. Leukemia & Lymphoma 43(8): 1589–1598.CrossRefGoogle Scholar
Ghossein, RA, Rosai, J (1996) Polymerase chain reaction in the detection of micrometastases and circulating tumor cells. Cancer 78(1): 10–16.3.0.CO;2-L>CrossRefGoogle ScholarPubMed
Dendukuri, N, Khetani, K, et al. (2007) Testing for HER2-positive breast cancer: a systematic review and cost-effectiveness analysis. CMAJ: Canadian Medical Association Journal 176(10): 1429–1434.CrossRefGoogle ScholarPubMed
Holodniy, M (1994) Clinical application of reverse transcription-polymerase chain reaction for HIV infection. Clinics in Laboratory Medicine 14(2): 335–349.Google ScholarPubMed
Kaplan, JC, Kahn, A, et al. (1992) Illegitimate transcription: its use in the study of inherited disease. Human Mutation 1(5): 357–360.CrossRefGoogle Scholar
Spitzack, KD, Ugaz, VM (2006) Polymerase chain reaction in miniaturized systems: big progress in little devices. Methods in Molecular Biology (Clifton, N.J.) 321: 97–129.Google ScholarPubMed
Rasmussen, A, Deckert, V (2005) New dimension in nano-imaging: breaking through the diffraction limit with scanning near-field optical microscopy. Analytical and Bioanalytical Chemistry 381(1): 165–172.CrossRefGoogle ScholarPubMed

Save book to Kindle

To save this book to your Kindle, first ensure [email protected] is added to your Approved Personal Document E-mail List under your Personal Document Settings on the Manage Your Content and Devices page of your Amazon account. Then enter the ‘name’ part of your Kindle email address below. Find out more about saving to your Kindle.

Note you can select to save to either the @free.kindle.com or @kindle.com variations. ‘@free.kindle.com’ emails are free but can only be saved to your device when it is connected to wi-fi. ‘@kindle.com’ emails can be delivered even when you are not connected to wi-fi, but note that service fees apply.

Find out more about the Kindle Personal Document Service.

Available formats
×

Save book to Dropbox

To save content items to your account, please confirm that you agree to abide by our usage policies. If this is the first time you use this feature, you will be asked to authorise Cambridge Core to connect with your account. Find out more about saving content to Dropbox.

Available formats
×

Save book to Google Drive

To save content items to your account, please confirm that you agree to abide by our usage policies. If this is the first time you use this feature, you will be asked to authorise Cambridge Core to connect with your account. Find out more about saving content to Google Drive.

Available formats
×