J Mol Diagn. 2016 Feb 24. pii: S1525-1578(16)00052-0. doi: 10.1016/j.jmoldx.2016.01.007. [Epub ahead of print]

The Application of Molecular Diagnostics to Stained Cytology Smears.

Oktay MH1, Adler E2, Hakima L2, Grunblatt E3, Pieri E3, Seymour A2, Khader S2, Cajigas A2, Suhrland M2, Goswami S4.

Author information

• 1Department of Pathology, Albert Einstein College of Medicine/Montefiore Medical Center, Bronx, New York. Electronic address: moktay@montefiore.org.
• 2Department of Pathology, Albert Einstein College of Medicine/Montefiore Medical Center, Bronx, New York.
• 3Department of Biology, Yeshiva University, New York, New York.
• 4Department of Biology, Yeshiva University, New York, New York. Electronic address: goswami@yu.edu.

Abstract

Detection of mutational alterations is important for guiding treatment decisions of lung non-small-cell carcinomas and thyroid nodules with atypical cytologic findings. Inoperable lung tumors requiring further testing for staging and thyroid lesions often are diagnosed using only cytology material.Molecular diagnostic tests of these samples typically are performed on cell blocks; however, insufficient cellularity of cell blocks is a limitation for test performance. In addition, some of the fixatives used while preparing cell blocks often introduces artifacts for mutation detection. Here, we applied qClamp xenonucleic technology and quantitative RT-PCR to cells microdissected directly from stained cytology smears to detect common alterations including mutations and translocations in non-small-cell carcinomas and thyroid lesions. By using this approach, we achieved a 1%molecular alteration detection rate from as few as 50 cells. Ultrasensitive methods of molecular alteration detection similar to the one described here will be increasingly important for the evaluation of molecular alterations in clinical scenarios when only small tissue samples are available.