Abstract
We present a protocol for the very rapid and sensitive detection of a specific mutation of the COL4A5 gene (exon 29, A-C mismatch) which was found in people affected by Alport syndrome (AS) and their families. Disposable electrochemically printed electrodes were used to immobilize a single-stranded oligonucleotide probe that was complementary to the AS-correlated gene. The detection principle is based on changes in the impedance spectra of the redox probe ferro/ferricyanide after hybridization with synthetic target DNA. Detection was performed either for mutated or for healthy (wild-type) gene copies. The high sensitivity obtained with this protocol (LOD in the picomolar range) was additionally enhanced to the femtomolar range by performing the detection in the presence of Ca2+. In fact, the specific binding of the metal ions in the presence of an A-C nucleotide mismatch induced a further impedance change, thus improving the discrimination between the mutated and healthy gene, as the signal amplification is achieved only for the former.
| Original language | English |
|---|---|
| Pages (from-to) | 3772-3779 |
| Number of pages | 8 |
| Journal | Analytical Chemistry |
| Volume | 82 |
| Issue number | 9 |
| DOIs | |
| Publication status | Published - 2010 May 1 |
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All Science Journal Classification (ASJC) codes
- Analytical Chemistry
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Rapid, sensitive, and label-free impedimetric detection of a single-nucleotide polymorphism correlated to kidney disease. / Bonanni, Alessandra; Pumera, Martin; Miyahara, Yuji.
In: Analytical Chemistry, Vol. 82, No. 9, 01.05.2010, p. 3772-3779.Research output: Contribution to journal › Article
TY - JOUR
T1 - Rapid, sensitive, and label-free impedimetric detection of a single-nucleotide polymorphism correlated to kidney disease
AU - Bonanni, Alessandra
AU - Pumera, Martin
AU - Miyahara, Yuji
PY - 2010/5/1
Y1 - 2010/5/1
N2 - We present a protocol for the very rapid and sensitive detection of a specific mutation of the COL4A5 gene (exon 29, A-C mismatch) which was found in people affected by Alport syndrome (AS) and their families. Disposable electrochemically printed electrodes were used to immobilize a single-stranded oligonucleotide probe that was complementary to the AS-correlated gene. The detection principle is based on changes in the impedance spectra of the redox probe ferro/ferricyanide after hybridization with synthetic target DNA. Detection was performed either for mutated or for healthy (wild-type) gene copies. The high sensitivity obtained with this protocol (LOD in the picomolar range) was additionally enhanced to the femtomolar range by performing the detection in the presence of Ca2+. In fact, the specific binding of the metal ions in the presence of an A-C nucleotide mismatch induced a further impedance change, thus improving the discrimination between the mutated and healthy gene, as the signal amplification is achieved only for the former.
AB - We present a protocol for the very rapid and sensitive detection of a specific mutation of the COL4A5 gene (exon 29, A-C mismatch) which was found in people affected by Alport syndrome (AS) and their families. Disposable electrochemically printed electrodes were used to immobilize a single-stranded oligonucleotide probe that was complementary to the AS-correlated gene. The detection principle is based on changes in the impedance spectra of the redox probe ferro/ferricyanide after hybridization with synthetic target DNA. Detection was performed either for mutated or for healthy (wild-type) gene copies. The high sensitivity obtained with this protocol (LOD in the picomolar range) was additionally enhanced to the femtomolar range by performing the detection in the presence of Ca2+. In fact, the specific binding of the metal ions in the presence of an A-C nucleotide mismatch induced a further impedance change, thus improving the discrimination between the mutated and healthy gene, as the signal amplification is achieved only for the former.
UR - http://www.scopus.com/inward/record.url?scp=77951770651&partnerID=8YFLogxK
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U2 - 10.1021/ac100165q
DO - 10.1021/ac100165q
M3 - Article
C2 - 20364839
AN - SCOPUS:77951770651
VL - 82
SP - 3772
EP - 3779
JO - Analytical Chemistry
JF - Analytical Chemistry
SN - 0003-2700
IS - 9
ER -