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Disadvantages

Use of RT-PCR for NTRK gene fusion testing requires that target sequences are known and the use of NTRK gene and exon-specific primers [1-3]. However, there are a relatively small number of NTRK genes and exons that are covered using this technique and RT-PCR uses RNA, which may not always be of sufficient quality due to its labile nature. Also, there is limited multiplexing ability as RT-PCR can only look at detect a few targets, whereas NGS can look at hundreds of targets at the same time.

 References

  1. Argani P, Fritsch M, Kadkol SS et al. Detection of the ETV6-NTRK3 chimeric RNA of infantile fibrosarcoma/cellular congenital mesoblastic nephroma in paraffin-embedded tissue: application to challenging pediatric renal stromal tumors. Mod Pathol 2000; 13: 29-36.
  2. Xu T, Wang H, Huang X et al. Gene Fusion in Malignant Glioma: An Emerging Target for Next-Generation Personalized Treatment. Transl Oncol 2018; 11: 609-618.
  3. Takeuchi K, Choi YL, Soda M et al. Multiplex reverse transcription-PCR screening for EML4-ALK fusion transcripts. Clin Cancer Res 2008; 14: 6618-6624.

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