Comparison of PNA clamping and direct sequencing for detecting KRAS mutations in matched tumour tissue, cell block, pleural effusion and serum from patients with malignant pleural effusion. Issue 1 (9th October 2014)

• Record Type: Journal Article
• Authors: Kang, Ji Young; Park, Chan Kwon; Yeo, Chang Dong; Lee, Hea Yeon; Rhee, Chin Kook; Kim, Seung Joon; Kim, Seok Chan; Kim, Young Kyoon; Park, Mi Sun; Yim, Hyeon Woo
• Is Part Of: Respirology. Volume 20:Issue 1(2015)
• Page Start: 138
• Page End: 146
• Publication Date: 2014-10-09
• Subjects: Respiratory organs -- Diseases -- Periodicals; Respiratory organs -- Periodicals; 612.2
• DOI: 10.1111/resp.12413
• Languages: English
• ISSNs: 1323-7799

Abstract

<abstract abstract-type="main"> <title>Abstract</title> <sec id="resp12413-sec-0001" sec-type="section"> <title>Background and objective</title> <p>Peptide nucleic acid (PNA)‐mediated real‐time polymerase chain reaction clamping was recently developed to improve mutation detection sensitivity. Pleural effusion could be a good sample candidate for mutation analysis. To establish if PNA clamping could be used to detect <italic>KRAS</italic> mutation in particular in pleural effusion, we analysed its diagnostic performance.</p> </sec> <sec id="resp12413-sec-0002" sec-type="section"> <title>Methods</title> <p>We studied 57 patients with malignant effusion. <italic>KRAS</italic> mutation was evaluated in samples of matched tumour tissue, cell block, pleural effusion and serum using PNA clamping and direct sequencing.</p> </sec> <sec id="resp12413-sec-0003" sec-type="section"> <title>Results</title> <p>The detection rate of <italic>KRAS</italic> mutation using pleural effusion was 14% for PNA clamping and 10.5% for direct sequencing. The κ coefficient between the two methods was 0.76 (<italic>P</italic> value < 0.0001), 1.00 (<italic>P</italic> value < 0.0001) and 0.87 (<italic>P</italic> value < 0.0001) in pleural effusion, tissue and cell block, respectively. The diagnostic performance of <italic>KRAS</italic> mutation detection from pleural effusion compared with the results obtained for all samples combined showed that the sensitivity, specificity, positive predictive value and negative predictive value were as follows: 89, 100, 100 and 98%, respectively for PNA clamping; 67, 100, 100 and 94%, respectively for directing sequencing.</p> </sec> <sec id="resp12413-sec-0004" sec-type="section"> <title>Conclusions</title> <p>The current study suggests that PNA clamping had a good concordance with direct sequencing for the detection of <italic>KRAS</italic> mutation in patients with malignant effusion. Furthermore, the good diagnostic performance obtained from pleural effusion samples provides evidence that pleural effusion can be a useful source for detecting <italic>KRAS</italic> mutation in a clinical setting, in which the collection of tumour tissues is challenging.</p> </sec> </abstract>