Analysis of Microsatellite Instability by Melting Peak Analysis with BAT26 and BATlS Specific 1m
(>97%, data not shown) show MSI in both BAT26 and BAT25 markers. Furthermore, it has been reported recently that BAT26 alone can specifically identify all
MSI-H tumors from mismatch repair gene mutation-positive CRC patients [14].
The complete micro satellite marker set [5, 6] should be used if no MSI is
detectable by BAT26 and BAT25 or only one BAT marker shows MSI, to avoid
missing a potential MSI positive tumor.
Due to the differently labeled hybridization probes, it is also possible to perform a duplex BAT26/BAT25 LightCycler PCR, but separate amplification seems
to be more robust {data not shown). As seen in our previous work [15], the range
of the temperature during the melting point measurement can obviously influence the T m values. However, there is no significant variation of T m values of different MSS tumors or normal tissues if the range of melting peak temperature is
kept constant for all assays. Since micro satellite melting peak analysis is achieved
within an hour of PCR setup, it provides an efficient tool for high-throughput MSI
screening analyses.
References
1. Thibodeau SN, Bren G, Schaid D (1993) Microsatellite instability in cancer of the proximal
colon. Science 260:81681-81689
2. Fishel R, Lescoe MK, Rao MR, Copeland NG, Jenkins NA, Garber J, Kane M, Kolodner R
(1993) The human mutator gene homolog MSH2 and its association with hereditary nonpolyposis colon cancer. Cell 75:1027-3108
3. Aaltonen LA, Peltomaki P, Mecklin JP, Jarvinen H, Jass JR, Green JS, Lynch HT, Watson P, Tallqvist G, Juhola M et al (1994) Replication errors in benign and malignant tumors from
hereditary nonpolyposis colorectal cancer patients. Cancer Res 54:1645-1648
4. Lynch HT, Smyrk TC (1998) Identifying hereditary nonpolyposis colorectal cancer. N Engl J
Med 338:1537-1538
5. Dietmaier W, Wallinger S, Bocker T, Kullmann F, Fishel R, Ruschoff J (1997) Diagnostic
microsatellite instability: definition and correlation with mismatch repair protein expression. Cancer Res 57:4749-4756
6. Boland CR, Thibodeau SN, Hamilton SR, Sidransky D, Eshleman JR, Burt RW, Meltzer SJ,
Rodriguez-Bigas MA, Fodde R, Ranzani GN, Srivastava S (1998) A National Cancer Institute
Workshop on micro satellite instability for cancer detection and familial predisposition:
development of international criteria for the determination of micro satellite instability in
colorectal cancer. Cancer Res 58:5248-5257
7. Schlegel J, Vogt T, Munkel K, Ruschoff J (1996) DNA fingerprinting of mammalian cell lines
using nonradioactive arbitrarily primed PCR (AP-PCR). Biotechniques 20:178-180
8. Mansfield DC, Brown AF, Green DK, Carothers AD, Morris SW, Evans HJ, Wright AF (1994)
Automation of genetic linkage analysis using fluorescent microsatellite markers. Genomics
24:225-233
9. Gyapay G, Ginot F, Nguyen S, VignalA, Weissenbach J (1996) Genotyping procedures in linkage papping. Methods 9:91-97
10. Papadopoulos N, Nicolaides NC, Wei YF, Ruben SM, Carter KC, Rosen CA, Haseltine WA,
Fleischmann RD, Fraser CM, Adams MD, Venter JC, Hamilton SR, Peterson GM, Watson P,
Lynch HAT, Peltomaki P, Mecklin JP, de la Chapelle A, Kinzler KW, Vogelstein B (1994) Mutation of a mutL homolog in hereditary colon cancer. Science 263:1625-1629
11. Hoang JM, Cottu PH, Thuille B, Salmon RJ, Thomas G, Hamelin R (1997) BAT-26, an indicator of the replication error phenotype in colorectal cancers and cell lines. Cancer Res
57:300-303
(>97%, data not shown) show MSI in both BAT26 and BAT25 markers. Furthermore, it has been reported recently that BAT26 alone can specifically identify all
MSI-H tumors from mismatch repair gene mutation-positive CRC patients [14].
The complete micro satellite marker set [5, 6] should be used if no MSI is
detectable by BAT26 and BAT25 or only one BAT marker shows MSI, to avoid
missing a potential MSI positive tumor.
Due to the differently labeled hybridization probes, it is also possible to perform a duplex BAT26/BAT25 LightCycler PCR, but separate amplification seems
to be more robust {data not shown). As seen in our previous work [15], the range
of the temperature during the melting point measurement can obviously influence the T m values. However, there is no significant variation of T m values of different MSS tumors or normal tissues if the range of melting peak temperature is
kept constant for all assays. Since micro satellite melting peak analysis is achieved
within an hour of PCR setup, it provides an efficient tool for high-throughput MSI
screening analyses.
References
1. Thibodeau SN, Bren G, Schaid D (1993) Microsatellite instability in cancer of the proximal
colon. Science 260:81681-81689
2. Fishel R, Lescoe MK, Rao MR, Copeland NG, Jenkins NA, Garber J, Kane M, Kolodner R
(1993) The human mutator gene homolog MSH2 and its association with hereditary nonpolyposis colon cancer. Cell 75:1027-3108
3. Aaltonen LA, Peltomaki P, Mecklin JP, Jarvinen H, Jass JR, Green JS, Lynch HT, Watson P, Tallqvist G, Juhola M et al (1994) Replication errors in benign and malignant tumors from
hereditary nonpolyposis colorectal cancer patients. Cancer Res 54:1645-1648
4. Lynch HT, Smyrk TC (1998) Identifying hereditary nonpolyposis colorectal cancer. N Engl J
Med 338:1537-1538
5. Dietmaier W, Wallinger S, Bocker T, Kullmann F, Fishel R, Ruschoff J (1997) Diagnostic
microsatellite instability: definition and correlation with mismatch repair protein expression. Cancer Res 57:4749-4756
6. Boland CR, Thibodeau SN, Hamilton SR, Sidransky D, Eshleman JR, Burt RW, Meltzer SJ,
Rodriguez-Bigas MA, Fodde R, Ranzani GN, Srivastava S (1998) A National Cancer Institute
Workshop on micro satellite instability for cancer detection and familial predisposition:
development of international criteria for the determination of micro satellite instability in
colorectal cancer. Cancer Res 58:5248-5257
7. Schlegel J, Vogt T, Munkel K, Ruschoff J (1996) DNA fingerprinting of mammalian cell lines
using nonradioactive arbitrarily primed PCR (AP-PCR). Biotechniques 20:178-180
8. Mansfield DC, Brown AF, Green DK, Carothers AD, Morris SW, Evans HJ, Wright AF (1994)
Automation of genetic linkage analysis using fluorescent microsatellite markers. Genomics
24:225-233
9. Gyapay G, Ginot F, Nguyen S, VignalA, Weissenbach J (1996) Genotyping procedures in linkage papping. Methods 9:91-97
10. Papadopoulos N, Nicolaides NC, Wei YF, Ruben SM, Carter KC, Rosen CA, Haseltine WA,
Fleischmann RD, Fraser CM, Adams MD, Venter JC, Hamilton SR, Peterson GM, Watson P,
Lynch HAT, Peltomaki P, Mecklin JP, de la Chapelle A, Kinzler KW, Vogelstein B (1994) Mutation of a mutL homolog in hereditary colon cancer. Science 263:1625-1629
11. Hoang JM, Cottu PH, Thuille B, Salmon RJ, Thomas G, Hamelin R (1997) BAT-26, an indicator of the replication error phenotype in colorectal cancers and cell lines. Cancer Res
57:300-303