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Clinical Chemistry 44: 1404-1409, 1998;
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(Clinical Chemistry. 1998;44:1404-1409.)
© 1998 American Association for Clinical Chemistry, Inc.

Molecular Diagnostics and Genetics

Validation of the Point-EXACCT method in non-small cell lung carcinomas

Veerle A. M. C. Somers1,a, Darcy A. Leimbach2, Paul H. M. H. Theunissen3, James J. Murtagh, Jr.2, Brian Holloway4, Anton W. Ambergen5, and Frederik B. J. M. Thunnissen1

1 Department of Pathology, Maastricht University, 6200 MD Maastricht, The Netherlands.

2 Pulmonary and Critical Care Medicine, Atlanta VA Medical Center, Emory University, Decatur, Georgia 30033.

3 Department of Pathology, De Wever Hospital, 6401 CX Heerlen, The Netherlands.

4 Centers for Disease Control and Prevention, National Center for Infectious Diseases, Atlanta, Georgia 30333.

5 Department of Methodology and Statistics, Maastricht University, 6200 MD Maastricht, The Netherlands.
a Address correspondence to this author at: Maastricht University, P.O. Box 616, 6200 MD Maastricht, The Netherlands. Fax 31-43-387-6613; e-mail VSO{at}lpat.azm.nl.

K-ras point mutations are often detected in part of the lung carcinomas. For the validation of a highly sensitive and rapid assay for known point mutations, Point-EXACCT (Biochim Biophys Acta 1998; 1379:42–52), we analyzed 89 non-small cell lung carcinomas and compared the results with two sequencing methods. No point mutations were found with double-stranded sequencing. Single-stranded sequencing detected six patients positive for K-ras codon 12. When Point-EXACCT was used, K-ras codon 12 mutations were detected in 8 of 52 patients with squamous cell carcinomas, 10 of 29 patients with adenocarcinomas, and 3 of 8 patients with large cell carcinomas. The finding of K-ras mutations in squamous cell carcinomas is explained by the high sensitivity of the method. Therefore, Point-EXACCT may be applicable to detection of those alterations occurring at a low frequency among an excess of cells with wild-type DNA.




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