Multiplexed genetic analysis using an expanded genetic alphabet.
Multiplexed genetic analysis using an expanded genetic alphabet.
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DOI：
10.1373/clinchem.2004.034330
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发表时间：
2004-11
影响因子：
9.3
通讯作者：
Prudent JR
中科院分区：
文献类型：
--
作者：
Johnson SC;Marshall DJ;Harms G;Miller CM;Sherrill CB;Beaty EL;Lederer SA;Roesch EB;Madsen G;Hoffman GL;Laessig RH;Kopish GJ;Baker MW;Benner SA;Farrell PM;Prudent JR
Background: All states require some kind of testing for newborns, but the policies are far from standardized. In some states, newborn screening may include genetic tests for a wide range of targets, but the costs and complexities of the newer genetic tests inhibit expansion of newborn screening. We describe the development and technical evaluation of a multiplex platform that may foster increased newborn genetic screening. Methods: MultiCode® PLx involves three major steps: PCR, target-specific extension, and liquid chip decoding. Each step is performed in the same reaction vessel, and the test is completed in ∼3 h. For site-specific labeling and room-temperature decoding, we use an additional base pair constructed from isoguanosine and isocytidine. We used the method to test for mutations within the cystic fibrosis transmembrane conductance regulator (CFTR) gene. The developed test was performed manually and by automated liquid handling. Initially, 225 samples with a range of genotypes were tested retrospectively with the method. A prospective study used samples from >400 newborns. Results: In the retrospective study, 99.1% of samples were correctly genotyped with no incorrect calls made. In the perspective study, 95% of the samples were correctly genotyped for all targets, and there were no incorrect calls. Conclusions: The unique genetic multiplexing platform was successfully able to test for 31 targets within the CFTR gene and provides accurate genotype assignments in a clinical setting.
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影响因子：
9.3
作者：
Moser, MJ;Marshall, DJ;Prudent, JR
通讯作者：
Prudent, JR
影响因子：
2.9
作者：
SWITZER, CY;MORONEY, SE;BENNER, SA
通讯作者：
BENNER, SA
影响因子：
5.6
作者：
Gerry, NP;Witowski, NE;Barany, F
通讯作者：
Barany, F
影响因子：
3
作者：
Day, NS;Tadin, M;Brown, S
通讯作者：
Brown, S
影响因子：
14.9
作者：
Johnson, SC;Sherrill, CB;Prudent, JR
通讯作者：
Prudent, JR