Detection of HER2 Status in Breast Cancer: Comparison of Current Methods with MLPA and Real-time RT-PCR

Pazhoomand, Reza; Keyhan, Elahe; Banan, Mehdi; Najmabad, Hossein; Karimlou, Masoud; Khodadad, Faranak; Iraniparast, Alireza; Feiz, Farnaz; Majidzadeh, Keivan; Bahman, Ideh; Moghadam, Fatemeh Aghakhani; Sobhani, Atoosa Madadkar; Abedin, Seyedeh Sedigheh; Muhammadnejad, Ahad; Behjat, Farkhondeh

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자료유형: 학술저널

저자정보: Pazhoomand, Reza (Genetics Research Center) Keyhan, Elahe (Genetics Research Center) Banan, Mehdi (Genetics Research Center) Najmabad, Hossein (Genetics Research Center) Karimlou, Masoud (Department of Biostatistics and Computer Sciences, University of Social Welfare and Rehabilitation Sciences) Khodadad, Faranak (Department of Surgery, Mehrad Hospital) Iraniparast, Alireza (School of Medicine, Shiraz University of Medical Sciences) Feiz, Farnaz (School of Medicine, Shiraz University of Medical Sciences) Majidzadeh, Keivan (Cancer Genetics Research Group, Breast Cancer Research Center, ACER) Bahman, Ideh (Genetics Research Center) Moghadam, Fatemeh Aghakhani (Genetics Research Center) Sobhani, Atoosa Madadkar (Department of Biostatistics and Computer Sciences, University of Social Welfare and Rehabilitation Sciences) Abedin, Seyedeh Sedigheh (Genetics Research Center) Muhammadnejad, Ahad (Cancer Research Center, Cancer Institute of Iran, Tehran University of Medical Sciences) Behjat, Farkhondeh (Genetics Research Center)

저널정보: 아시아태평양암예방학회 Asian Pacific journal of cancer prevention : APJCP Asian Pacific journal of cancer prevention : APJCP 제14권 제12호

발행연도: 2013.1

수록면: 7,621 - 7,628 (8page)

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Human epidermal growth factor receptor (HER) status is an important prognostic factor in breast cancer. There is no globally accepted method for determining its status, and which method is most precise is still a matter of debate. We here analyzed HER2 mRNA expression by quantitative reverse transcription-PCR (qRT-PCR) and HER2 DNA amplification using multiplex ligation-dependent probe amplification (MLPA). In parallel, we performed a routine evaluation of HER2 protein by immunohistochemistry (IHC). To assess the accuracy of the RT-PCR and MLPA techniques, a combination of IHC and fluorescence in situ hybridization (FISH) was used, substituting FISH when the results of IHC were ambiguous (2+) and for those IHC results that disagreed with MLPA and qRT-PCR, this approach being termed IHC-FISH. The IHC results for four samples were not compatible with the MLPA and qRT-PCR results; the MLPA and qRT-PCR results for these samples were confirmed by FISH. The correlations between IHC-FISH and qRT-PCR or MLPA were 0.945 and 0.973, respectively. The ASCO/CAP guideline IHC/FISH correlation with MLPA was (0.827) and with RT-PCR was (0.854). The correlations between the IHC results (0, 1+ as negative, and 3+ as positive) and qRT-PCR and MLPA techniques were 0.743 and 0.831, respectively. Given the shortcomings of IHC analysis and greater correlations between MLPA, qRT-PCR, and FISH methods than IHC analysis alone with each of these three methods, we propose that MLPA and real-time PCR are good alternatives to IHC. However a suitable cut-off point for qRTPCR is a prerequisite for determining the exact status of HER2.

#Breast cancer #HER2 status #MLPA #IHC #FISH #RT-PCR #method comparison

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