Table 1

Advantages and disadvantages of the major techniques applied on cytological material

AuthorEGFR detectionKRAS detectionCytological materialType of fixationAdvantagesDisadvantages
Janne et al, 200625SURVEYOR digestion, RFLP assay and dsDNA sizing by high-performance liquid chromatographyNACell lines and lung biopsiesFrozen and FFPEDoes not require microdissection and more sensitive than the direct sequencing methodLaborious and expensive
Nomoto et al, 200629HRMANAFNAEthanol fixedHigh sensitivity and specificityDetection of known mutations
Smith et al, 200731HRMANAFNAAir-driedHigh sensitivity and specificityDetection of known mutations
Molina et al, 200836Laser microdissection in PCR buffer and detection by length analysis of fluorescently labelled PCR products and TaqMan assayNACell lines on slideFresh and FFPENo need to DNA purification and enrichment of tumour cellsTime consuming, laborious and expensive
Lim et al, 200937WGA and direct sequencingWGA and direct sequencingLung biopsies and FNARNA Later (Ambion)Useful to expand DNA from low-volume biopsiesRequires time and an appropriate WGA kit
Wu et al, 200838Reverse transcriptase PCR coupled direct sequencingNAPleural effusionsTri-reagent (Molecular Research Center)Useful to predict the mutational status in pleural effusionsLimited sensitivity, high cost, and time consuming
Kimura et al, 200639Scorpion amplified refractory mutation systemNAPleural effusionsCell pellets frozen at −80°CMore sensitive than the direct sequencing method, simple and very fastThe EGFR Scorpion kit detects the two major somatic mutations
Wei et al, 200740FISHNABronchial washings and FNAFFPEFISH is useful for detecting chromosomal abnormalitiesUnable to detect gene mutations
Yatabe et al, 200641Reverse transcriptase PCR-coupled direct sequencingNASmall biopsiesFFPEDoes not require a tumour cell enrichment procedureLimited sensitivity, high cost, and time consuming
Horiike et al, 200742Scorpion amplified refractory mutation systemNAFNAStore at −80°C in saline solutionMore sensitive than the direct sequencing methodThe EGFR Scorpion kit detects the two major somatic mutations
  • NA, not analysed; HRMA, high resolution melting analysis; WGA, whole genome amplification; FNA, fine needle aspirates; FFPE, formalin fixed paraffin embedded; FISH, fluorescence in situ hybridisation; RFLP, restriction fragment length polymorphism.