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Scientia Pharmaceutica is published by MDPI from Volume 84 Issue 3 (2016). Previous articles were published by another publisher in Open Access under a CC-BY (or CC-BY-NC-ND) licence, and they are hosted by MDPI on as a courtesy and upon agreement with Austrian Pharmaceutical Society (Österreichische Pharmazeutische Gesellschaft, ÖPhG).
Correction published on 2 December 2016, see Sci. Pharm. 2016, 84(4), 753.

Detection of HER2 Gene Polymorphism in Breast Cancer: PCR Optimization Study

Research Center for Biotechnology, Indonesian Institute of Sciences (LIPI), Jalan Raya Bogor Km. 46, Cibinong 16911, Indonesia.
Author to whom correspondence should be addressed.
Sci. Pharm. 2016, 84(1), 103-111;
Received: 5 September 2015 / Accepted: 15 December 2015 / Published: 14 February 2016
Cancers are the most deadly diseases in the world and their incidences continue to increase over time. Particularly, breast cancer in females places 1st rank among other types of cancers in term of cancer cases (23%) and death incidence (14%). Recent findings support the correlation between Ile655Val SNP in the HER2 gene with breast cancer risk. Moreover, the Ile655Val HER2 gene polymorphism could be a predictive factor in a neoadjuvant therapy setting. Precise detection of the Ile655Val HER2 gene SNP in early breast cancer patients will be beneficial in designing the most suitable treatment and in increasing the efficacy of anticancer drugs. Here we develop a rapid and inexpensive method for Ile655Val SNP detection in the HER2 gene based on allele-specific PCR technology. Two forward primers and one common reverse primer were designed to anneal specifically either on the HER2 gene fragment containing the GG genotype or to the HER2 gene fragment containing the AA genotype where one of these primers had been added with poly-GC at 5’ upstream. Moreover, to increase discrimination level, mismatch bases at the SNP site and the 3rd base of each forward primers from 3’end were added. To test the performance of the designed primers in discriminating a polymorphism and its annealing temperature, breast cancer specimen-derived genomic DNA (with GG genotype) and pGEM_HER2/AA (with AA genotype) were used as templates in the PCR reaction. The optimal annealing temperature for SNP detection was at 51.5°C as showed by the appearance of a 150 base pair (bp) band as AA genotype (pGEM_HER2/AA template), 116bp band as GG genotype (genomic DNA template), and both types of bands as AG genotype (mix of pGEM_HER2/AA and genomic DNA template). Allelic types of breast cancer patients were also determined using this optimized method compared to sanger sequencing. The 100% accordance was shown for all types of genotypes in both methods. The allele-specific PCR in this study may have application in determining polymorphisms of the breast cancers-originated Ile655Val HER2 gene.
Keywords: Breast cancer; HER2 gene; SNP; Allele-Specific PCR Breast cancer; HER2 gene; SNP; Allele-Specific PCR
MDPI and ACS Style

BUDIARTO, B.R.; DESRIANI. Detection of HER2 Gene Polymorphism in Breast Cancer: PCR Optimization Study. Sci. Pharm. 2016, 84, 103-111.

AMA Style

BUDIARTO BR, DESRIANI. Detection of HER2 Gene Polymorphism in Breast Cancer: PCR Optimization Study. Scientia Pharmaceutica. 2016; 84(1):103-111.

Chicago/Turabian Style

BUDIARTO, Bugi R., and DESRIANI. 2016. "Detection of HER2 Gene Polymorphism in Breast Cancer: PCR Optimization Study" Scientia Pharmaceutica 84, no. 1: 103-111.

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