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Microsatellite instability test using peptide nucleic acid probe-mediated melting point analysis: a comparison study

 Mi Jang  ;  Yujin Kwon  ;  Hoguen Kim  ;  Hyunki Kim  ;  Byung Soh Min  ;  Yehyun Park  ;  Tae Il Kim  ;  Sung Pil Hong  ;  Won Kyu Kim 
 BMC CANCER, Vol.18(1) : 1218, 2018 
Journal Title
Issue Date
Colorectal cancer ; Microsatellite instability ; Peptide nucleic acid probe and immunohistochemistry ; Real-time polymerase chain reaction
BACKGROUND: Analysis of high microsatellite instability (MSI-H) phenotype in colorectal carcinoma (CRC) is important for evaluating prognosis and choosing a proper adjuvant therapy. Although the conventional MSI analysis methods such as polymerase chain reaction (PCR) fragment analysis and immunohistochemistry (IHC) show high specificity and sensitivity, there are substantial barriers to their use.

METHODS: In this study, we analyzed the MSI detection performance of three molecular tests and IHC. For the molecular tests, we included a recently developed peptide nucleic acid probe (PNA)-mediated real-time PCR-based method using five quasi-monomorphic mononucleotide repeat markers (PNA method) and two conventional PCR fragment analysis methods using NCI markers (NCI method) or five quasi-monomorphic mononucleotide repeat markers (MNR method). IHC analysis was performed with four mismatch repair proteins. The performance of each method was validated in 166 CRC patient samples, which consisted of 76 MSI-H and 90 microsatellite stable (MSS) CRCs previously diagnosed by NCI method.

RESULTS: Of the 166 CRCs, 76 MSI-H and 90 MSS CRCs were determined by PNA method. On the other hand, 75 MSI-H and 91 MSS CRCs were commonly determined by IHC and MNR methods. Based on the originally diagnosed MSI status, PNA showed 100% sensitivity and 100% specificity while IHC and MNR showed 98.68% sensitivity and 100% specificity. When we analyzed the maximum sensitivity of MNR and PNA method, which used the same five markers, PNA method could detect alterations in all five mononucleotide repeat markers in samples containing down to 5% MSI-H DNAs, whereas MNR required at least 20% MSI-H DNAs to achieve the same performance.

CONCLUSIONS: Based on these findings, we suggest that PNA method can be used as a practical laboratory test for the diagnosis of MSI.
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1. College of Medicine (의과대학) > Dept. of Internal Medicine (내과학교실) > 1. Journal Papers
1. College of Medicine (의과대학) > Dept. of Pathology (병리학교실) > 1. Journal Papers
1. College of Medicine (의과대학) > Dept. of Surgery (외과학교실) > 1. Journal Papers
1. College of Medicine (의과대학) > Yonsei Biomedical Research Center (연세의생명연구원) > 1. Journal Papers
Yonsei Authors
Kim, Won Kyu(김원규)
Kim, Tae Il(김태일) ORCID logo https://orcid.org/0000-0003-4807-890X
Kim, Hyunki(김현기) ORCID logo https://orcid.org/0000-0003-2292-5584
Kim, Hogeun(김호근)
Min, Byung Soh(민병소) ORCID logo https://orcid.org/0000-0003-0180-8565
Park, Yehyun(박예현) ORCID logo https://orcid.org/0000-0001-8811-0631
Jang, Mi(장미) ORCID logo https://orcid.org/0000-0002-0153-9847
Hong, Sung Pil(홍성필)
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