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Microsatellite instability typing in serum and tissue of patients with colorectal cancer: comparing real time PCR with hybridization probe and high-performance liquid chromatography

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Abstract

Allelic variation of BAT-25 (a 25-repeat quasimonomorphic poly T) and BAT-26 (a 26-repeat quasimonomorphic polyA) loci as two mononucleotide microsatellite markers, were analyzed with high-performance liquid chromatography (HPLC) compared with Real-Time PCR using hybridization probes. BAT-26 and BAT-25 markers were used to determine an appropriate screening technique with high sensitivity and specificity to diagnose microsatellite instability (MSI) status in patients with colorectal cancer (CRC). One of the pathways in colorectal tumor genesis is microsatellite instability (MSI+). MSI is detected in about 15 % of all CRCs; 3 % are of these are associated with Lynch syndrome and the other 12 % are caused by sporadic. Colorectal tumors with MSI have distinctive features compared with microsatellite stable tumors. Due to the high percentage of MSI+ CRC in Iran, screening of this type of CRC is imperative. Two markers were analyzed in tissues and sera of 44 normal volunteers and tumor and matched normal mucosal tissues as well as sera of 44 patients with sporadic CRC. The sensitivity and specificity of BAT-26 with real time PCR method (Hybridization probe) were 100 % in comparison with sequencing method as the gold standard, while HPLC had a lower sensitivity and specificity. According to HPLC data, BAT-26 was more sensitive than BAT-25 in identifying MSI tumors. Therefore, MSI typing using the BAT-26 hybridization probe method compared to HPLC could be considered as an accurate method for diagnosing MSI in CRC tumors but not in serum circulating DNAs.

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Acknowledgments

The present Article was extracted from the thesis written by Marjan Rismanchi and financially supported by Shiraz University of Medical Sciences Grant No. 90-5532.

Conflict of interest

The authors declared no conflict of interest.

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Authors and Affiliations

  1. Department of Biochemistry, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran

    P. Mokarram, Z. Honardar, S. Kavousipour, F. Naghibalhossaini & Z. Mostafavi-Pour

  2. Department of Biochemistry, School of Medicine, Student Research Committee, Shiraz University of Medical Sciences, Shiraz, Iran

    M. Rismanchi

  3. Gastroenterohepatology Research Center, Nemazee Hospital, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran

    P. Mokarram, M. Alizadeh Naeeni & S. A. Shamsdin

  4. Department of Internal Medicine, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran

    M. Alizadeh Naeeni

  5. Food and Drug Laboratory Research Center, Ministry of Health and Medical Education, Tehran, Iran

    S. Mirab Samiee

  6. Department of Molecular Pathology, Day General Hospital Laboratory, Tehran, Iran

    S. Mirab Samiee

  7. Research and Development Department, Production and Research Complex, Pasteur Institute of Iran, Tehran, Iran

    M. Paryan

  8. Community Medicine Department, Medical Faculty, Mazandaran University of Medical Sciences, Sari, Iran

    A. Alipour

  9. Department of Pathology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran

    A. Monabati

  10. Colorectal Research Center, Shiraz University of Medical Sciences, Shiraz, Iran

    P. Mokarram & S. V. Hosseni

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  1. P. Mokarram
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  2. M. Rismanchi
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  9. F. Naghibalhossaini
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  10. Z. Mostafavi-Pour
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Correspondence to P. Mokarram.

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Mokarram, P., Rismanchi, M., Alizadeh Naeeni, M. et al. Microsatellite instability typing in serum and tissue of patients with colorectal cancer: comparing real time PCR with hybridization probe and high-performance liquid chromatography. Mol Biol Rep 41, 2835–2844 (2014). https://doi.org/10.1007/s11033-014-3138-1

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  • DOI: https://doi.org/10.1007/s11033-014-3138-1

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