With the rapid expansion of genomic health care, nurses are exposed to emerging genetic technologies in a wide variety of clinical and research settings; however, nurses have limited knowledge about these technologies. The polymerase chain reaction procedure, which is the foundation of current molecular genetic technologies, real-time polymerase chain reaction, and microarray analysis are described in this article. The applications, strengths, and limitations of each technology are discussed.
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References
1.
Abe A, Inoue K, Tanaka T, Kato J, Kajiyama N, Kawaguchi R, Tanaka S, Yoshiba M, Kohara M. 1999. Quantitation of hepatitis B virus genomic DNA by real-time detection PCR. J Clin Microbiol37:2899-2903.
2.
Abravaya K, Huff J, Marshall R, Merchant B, Mullen C, Schneider G, Robinson J. 2003. Molecular beacons as diagnostic tools: technology and applications. Clin Chem Lab Med41:468-474.
Afshari CA. 2002. Perspective: microarray technology, seeing more than spots. Endocrinology143:1983-1989.
5.
Ballerini S, Bellincampi L, Bernardini S, Casciani S, Motti C, Cortese C, Federici G. 2002. Apolipoprotein E genotyping: a comparative study between restriction endonuclease mapping and allelic discrimination with the LightCycler. Clin Chim Acta317:71-76.
6.
Barton MC, Stivers DN. 2002. Microarray analysis of hepaticregulated gene expression: specific applications and nonspecific problems. Hepatology35:727-729.
7.
Bernard PS, Wittwer CT. 2002. Real-time PCR technology for cancer diagnostics. Clin Chem48:1178-1185.
8.
Bieche I, Onody P, Laurendeau I, Olivi M, Vidaud D, Lidereau R, Vidaud M. 1999. Real-time reverse transcription-PCR assay for future management of ERBB2-based clinical applications. Clin Chem45:1148-1156.
9.
Bubendorf L, Nocito A, Moch H, Sauter G. 2001. Tissue microarray (TMA) technology: miniaturized pathology archives for high-throughput in situ studies. J Pathol195:72-79.
10.
Bustin SA. 2000. Absolute quantification of mRNA using real-time reverse transcription polymerase chain reaction assays. J Mol Endocrinol25:169-193.
11.
DePrimo SE, Wong LM, Khatry DB, Nicholas SL, Manning WC, Smolich BD, O’Farrell AM, Cherrington JM. 2003. Expression profiling of blood samples from an SU5416 phase III meta-static colorectal cancer clinical trial: a novel strategy for biomarker identification. BMC Cancer3:3.
12.
Dyrskjot L, Thykjaer T, Kruhoffer M, Jensen JL, Marcussen N, Hamilton-Dutoit S, Wolf H, Orntoft TF. 2003. Identifying distinct classes of bladder carcinoma using microarrays. Nat Genet33:90-96.
13.
Gibson UE, Heid CA, Williams PM. 1996. A novel method for real time quantitative RT-PCR. Genome Res6:995-1001.
14.
Greenberg DA. 2001. Microarrays, markers of disease, and the myth of “nonhypothesis-driven” research. Ann Neurol50:695.
15.
Hampton GM, Frierson HF. 2003. Classifying human cancer by analysis of gene expression. Trends Mol Med9:5-10.
16.
Hedenfalk I, Ringner M, Ben-Dor A, Yakhini Z, Chen Y, Chebil G, Ach R, Loman N, Olsson H, Meltzer P, and others. 2003. Molecular classification of familial non-BRCA1/BRCA2 breast cancer. Proc Natl Acad Sci U S A100:2532-2537.
17.
Heid CA, Stevens J, Livak KJ, Williams PM. 1996. Real time quantitative PCR. Genome Res6:986-994.
18.
Higgins JP, Shinghal R, Gill H, Reese JH, Terris M, Cohen RJ, Fero M, Pollack JR, Van De Rijn M, Brooks JD. 2003. Gene expression patterns in renal cell carcinoma assessed by complementary DNA microarray. Am J Pathol162:925-932.
19.
Higuchi R, Dollinger G, Walsh PS, Griffith R. 1992. Simultaneous amplification and detection of specific DNA sequences. Bio-technology (N Y)10:413-417.
20.
Holland PM, Abramson RD, Watson R, Gelfand DH. 1991. Detection of specific polymerase chain reaction product by utilizing the 5′→3′ exonuclease activity of Thermus aquaticus DNA polymerase. Proc Natl Acad Sci U S A88:7276-7280.
21.
Jenkins JF, Prows C, Dimond E, Monsen R, Williams J. 2001. Recommendations for educating nurses in genetics. J Prof Nurs17:283-290.
22.
Koshland DE Jr. 1989. The molecule of the year. Science246:1541.
23.
Li H, Dummer JS, Estes WR, Meng S, Wright PF, Tang YW. 2003. Measurement of human cytomegalovirus loads by quantitative real-time PCR for monitoring clinical intervention in transplant recipients. J Clin Microbiol41:187-191.
24.
Livak KJ, Schmittgen TD. 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods25:402-408.
25.
Maughan NJ, Lewis FA, Smith V. 2001. An introduction to arrays. J Pathol195:3-6.
26.
Mhlanga MM, Malmberg L. 2001. Using molecular beacons to detect single-nucleotide polymorphisms with real-time PCR. Methods25:463-471.
27.
Mullis KB. 1990. The unusual origin of the polymerase chain reaction. Sci Am262:56-61, 64-65.
28.
Nussbaum R, McInnes RR, Willard HF, Boerkoel CF. 2001. Thompson & Thompson genetics in medicine. Philadelphia: W. B. Saunders.
29.
Oliver DH, Thompson RE, Griffin CA, Eshleman JR. 2000. Use of single nucleotide polymorphisms (SNP) and real-time polymerase chain reaction for bone marrow engraftment analysis. J Mol Diagn2:202-208.
30.
Puhakka T, Lehti H, Vainionpaa R, Jormanainen V, Pulkkinen M, Sharp S, Kerr C, Dempsey M, Ring CJ, Ward C, and others. 2003. Zanamivir: a significant reduction in viral load during treatment in military conscripts with influenza. Scand J Infect Dis35:52-58.
31.
Sabek O, Dorak MT, Kotb M, Gaber AO, Gaber L. 2002. Quantitative detection of T-cell activation markers by real-time PCR in renal transplant rejection and correlation with histopathologic evaluation. Transplantation74:701-707.
32.
Sambrook J, Russell DW, editors. 2001a. Amplification of cDNA generated by reverse transcription of mRNA. In: Molecular cloning: a laboratory manual. 3rd ed. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press. p 8.46-8.53.
33.
Sambrook J, Russell DW, editors. 2001b. The basic polymerase chain reaction. In: Molecular cloning: a laboratory manual. 3rd ed. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press. p 8.18-8.24.
34.
Sanchez-Carbayo M. 2003. Use of high-throughput DNA microarrays to identify biomarkers for bladder cancer. Clin Chem49:23-31.
35.
Scherer A, Krause A, Walker JR, Korn A, Niese D, Raulf F. 2003. Early prognosis of the development of renal chronic allograft rejection by gene expression profiling of human protocol biopsies. Transplantation75:1323-1330.
36.
Shackel NA, Gorrell MD, McCaughan GW. 2002. Gene array analysis and the liver. Hepatology36:1313-1325.
37.
Shi L. 2002. DNA Microarray (genome chip)—monitoring the genome on a chip. Available from: http://www.genechips.com. Accessed on 2003 June 4.
38.
Smith MW, Yue ZN, Geiss GK, Sadovnikova NY, Carter VS, Boix L, Lazaro CA, Rosenberg GB, Bumgarner RE, Fausto N, and others. 2003. Identification of novel tumor markers in hepatitis C virus-associated hepatocellular carcinoma. Cancer Res63:859-864.
39.
Song P, Li S, Meibohm B, Gaber AO, Honaker MR, Kotb M, Yates CR. 2002. Detection of MDR1 single nucleotide polymorphisms C3435T and G2677T using real-time polymerase chain reaction: MDR1 single nucleotide polymorphism genotyping assay. AAPS PharmSci4:29.
40.
Soulet D, Rivest S. 2002. Perspective: how to make microarray, serial analysis of gene expression, and proteomic relevant to day-to-day endocrine problems and physiological systems. Endocrinology143:1995-2001.
41.
Tefferi A, Bolander ME, Ansell SM, Wieben ED, Spelsberg TC. 2002. Primer on medical genomics. Part III: microarray experiments and data analysis. Mayo Clin Proc77:927-940.
42.
Vernon SD, Unger ER, Dimulescu IM, Rajeevan M, Reeves WC. 2002. Utility of the blood for gene expression profiling and biomarker discovery in chronic fatigue syndrome. Dis Markers18:193-199.
43.
Wang D, Coscoy L, Zylberberg M, Avila PC, Boushey HA, Ganem D, DeRisi JL. 2002. Microarray-based detection and genotyping of viral pathogens. Proc Natl Acad Sci USA99:15687-15692.
44.
White PA, Pan Y, Freeman AJ, Marinos G, Ffrench RA, Lloyd AR, Rawlinson WD. 2002. Quantification of hepatitis C virus in human liver and serum samples by using LightCycler reverse transcriptase PCR. J Clin Microbiol40:4346-4348.
45.
Wu TD. 2001. Analysing gene expression data from DNA microarrays to identify candidate genes. J Pathol195:53-65.
46.
Yeoh EJ, Ross ME, Shurtleff SA, Williams WK, Patel D, Mahfouz R, Behm FG, Raimondi SC, Relling MV, Patel A, and others. 2002. Classification, subtype discovery, and prediction of out-come in pediatric acute lymphoblastic leukemia by gene expression profiling. Cancer Cell1:133-143.