Ra?id Detection of Missense Mutations in the Prostatic Steroid Sa-Reductase Gene 1m
Table 2. Melting temperatures of SRD5A2
Locus
Allele
Pairing
T m (0C) observed
SRD5A2; codon 49
49 alanine
G- C match
62.5
49 valine
A-C mismatch
51.0
SRD5A2; codon 89
89 leucine
C- Gmatch
57.5
89 valine
C- C mismatch
49.0
• The processing of the samples is simple and the analysis is fast, providing rapid results as well as the possibility of high throughput genotyping.
• As this method is performed in a closed system with no post-amplification
processing, potential problems with sample tracking and end-product contamination are eliminated.
• As hands-on time is shorter than in any other technique used so far and costs
for reagents and consumables are not higher than for conventional assays, the
fluorescence method allows SRD5A2 genotyping in a very economic manner.
• The simultaneous analysis of two polymorphisms by utilizing two different
reporter dyes and the color compensation software additionally saves time and
money.
In summary, this homogeneous (closed-tube) assay for rapid genotyping of the
SRD5A2 polymorphisms at co dons 49 and 89 on the LightCycler is ideallyapplicable to routine analysis in a clinical and/or research setting. This method may
facilitate further research on the association between polymorphisms in the
SRD5A2 gene and the risk of developing prostate cancer and eventually, when a
predictive value has been established, it may be used in the risk stratification of
patients and for selection of therapeutic strategies.
References
1. Ross RK, Pike MC, Coetzee GA, Reichardt JK, Yu MC, Feigelson H, Stanczyk FZ, Kolonel LN,
Henderson BE (1998) Androgen metabolism and prostate cancer: establishing a model of
genetic susceptibility. Cancer Res 58:4497-4504
2. Ross RK, Bernstein L, Lobo RA, Shimizu H, Stanczyk FZ, Pike MC, Henderson BE (1992) 5alpha-reductase activity and risk of prostate cancer among Japanese and US white and black
males. Lancet 339:887-889
3. Makridakis NM, Ross RK, Pike MC, Crocitto LE, Kolonel LN, Pearce CL, Henderson BE,
Reichardt JK (1999) Association of mis-sense substitution in SRDSA2 gene with prostate
cancer in African-American and Hispanic men in Los Angeles, USA. Lancet 354:975-978
4. Makridakis N, Ross RK, Pike MC, Chang L, Stanczyk FZ, Kolonel LN, Shi CY, Yu MC, Henderson BE, Reichardt JK (1997) A prevalent missense substitution that modulates activity of
prostatic steroid 5alpha-reductase. Cancer Res 57:1020-1022
Table 2. Melting temperatures of SRD5A2
Locus
Allele
Pairing
T m (0C) observed
SRD5A2; codon 49
49 alanine
G- C match
62.5
49 valine
A-C mismatch
51.0
SRD5A2; codon 89
89 leucine
C- Gmatch
57.5
89 valine
C- C mismatch
49.0
• The processing of the samples is simple and the analysis is fast, providing rapid results as well as the possibility of high throughput genotyping.
• As this method is performed in a closed system with no post-amplification
processing, potential problems with sample tracking and end-product contamination are eliminated.
• As hands-on time is shorter than in any other technique used so far and costs
for reagents and consumables are not higher than for conventional assays, the
fluorescence method allows SRD5A2 genotyping in a very economic manner.
• The simultaneous analysis of two polymorphisms by utilizing two different
reporter dyes and the color compensation software additionally saves time and
money.
In summary, this homogeneous (closed-tube) assay for rapid genotyping of the
SRD5A2 polymorphisms at co dons 49 and 89 on the LightCycler is ideallyapplicable to routine analysis in a clinical and/or research setting. This method may
facilitate further research on the association between polymorphisms in the
SRD5A2 gene and the risk of developing prostate cancer and eventually, when a
predictive value has been established, it may be used in the risk stratification of
patients and for selection of therapeutic strategies.
References
1. Ross RK, Pike MC, Coetzee GA, Reichardt JK, Yu MC, Feigelson H, Stanczyk FZ, Kolonel LN,
Henderson BE (1998) Androgen metabolism and prostate cancer: establishing a model of
genetic susceptibility. Cancer Res 58:4497-4504
2. Ross RK, Bernstein L, Lobo RA, Shimizu H, Stanczyk FZ, Pike MC, Henderson BE (1992) 5alpha-reductase activity and risk of prostate cancer among Japanese and US white and black
males. Lancet 339:887-889
3. Makridakis NM, Ross RK, Pike MC, Crocitto LE, Kolonel LN, Pearce CL, Henderson BE,
Reichardt JK (1999) Association of mis-sense substitution in SRDSA2 gene with prostate
cancer in African-American and Hispanic men in Los Angeles, USA. Lancet 354:975-978
4. Makridakis N, Ross RK, Pike MC, Chang L, Stanczyk FZ, Kolonel LN, Shi CY, Yu MC, Henderson BE, Reichardt JK (1997) A prevalent missense substitution that modulates activity of
prostatic steroid 5alpha-reductase. Cancer Res 57:1020-1022