Realtime quantification of HER2/neu gene amplification by polymerase chain reaction (PCR)

Beyser, Kurt; Reiser, Astrid; Gross, Cynthia; Stiefken, J.; Gloeckner-Hofmann, K.; Tabiti, Karim; Pedrocchi, M.; Rueschoff, Josef

doi:10.1016/s0959-8049(01)81144-8

Cited by 2 publications

(2 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…HER-2 quantification detection kit is already commercialized, such as by Roche. The kit provides specific primers, hybridization probes, positive and negative control reaction (Beyser et al 2001). Since the probes frequently used for the DNA labeling process hence it directly causes the kits price becoming slightly expensive.…”

Section: Pgem-t Easy Her-2 For Standard Curve Calculation In Scoring mentioning

confidence: 99%

Data provision of PIK3CA gene diversity and recombinant plasmids preparation for control DNA in developing the trastuzumab predictive response diagnostic kit

et al. 2016

View full text Add to dashboard Cite

HER-2 overexpression is well known as a poor prognostic factor for breast cancer patients. Targeted therapy can be carried out using monoclonal antibody named trastuzumab. Some reports have higlighted the core problem of HER2 positive-breast cancer resitance on trastuzumab due to incorrect in selecting HER2 status patient who will reciept the drug and the emergence of PIK3CA mutations especially in exon 9 and 20 which is the downstream of HER-2 pathway. In this study, data provision of PIK3CA gene and preparation of plasmid to support developing the trastuzumab predictive response diagnostic kits will be reported. Based on direct DNA sequencing result, two samples of 68 breast cancer patients exhibited mutation at exon 20 H1047R, while another three samples showed silent mutation (T1025T) at the same exon. On the other hand, careful strategy should be considered for exon 9 analysis, since we found that almost 68 samples sequenced none of them were exon 9 positive (pesudogene). Two prepared plasmids, pGEMT-easy PIK3CA exon 9 and 20 will be applied as control PIK3CA gene for qPCR SYBR green I-based PIK3CA genotyping, while pGEMT easy HER-2 will be applied as a reference gene for scoring HER-2 status. The standard curve equation of plasmid-cloned HER2 gene amplification was Y=-3,0472x + 46,465, R 2 = 0,99 with qPCR efficiency was 115%, respectively. Inconclusion, data provision and control DNA preparation of predicted factors for breast cancer patients who positively respond to trastuzumab are very fundamental important aspects for the development of trastuzumab response diagnostic kit which is based on Indonesian population genetics profile.

show abstract

Section: Pgem-t Easy Her-2 For Standard Curve Calculation In Scoring mentioning

confidence: 99%

Data provision of PIK3CA gene diversity and recombinant plasmids preparation for control DNA in developing the trastuzumab predictive response diagnostic kit

et al. 2016

View full text Add to dashboard Cite

show abstract

“…Polysomy occurred at 13% to 46% in breast cancer. Since there was a polysomic problem in chromosome 17 that potentially contributed to misinterpretation of HER-2 scoring status, He et al (2016) (Beyser et al 2001, Benohr et al 2005Murad et al 2013). Further Mendoza et al (2013), standardized qPCR analysis by using whn that localized also on chromosome 17.…”

Section: Introductionmentioning

confidence: 99%

Housekeeping gene as a source of calibrator candidate for HER-2 scoring in frozen tissue breast cancer study based on qPCR

et al. 2017

View full text Add to dashboard Cite

Abstract. Rismaya, Azamris, Budiarti S, Desriani. 2017 Here in this report, we investigated new reference gene coming from housekeeping gene as an alternative calibrator for HER-2 amplification scoring based on qPCR methods. We compared two recommended housekeeping gene, 18S rRNA and β-Actin, as a candidate for a HER-2 calibrator. For methods development, optimization and comparison study between 18S rRNA and β-Actin, we used non-invasive samples (DNA buccal cells). Selected candidate reference further was tested to eighteen breast cancer sample, validated with qPCR methods refer to Mendoza et al. 2013. β-Actin is shown as the best candidate for HER-2 scoring status determination. qPCR concordance result with other qPCR methods refers to Mendoza et al. 2013 shown 94.4%. The qPCR efficiency and % CV value shown as a requirement as a theory. qPCR efficiency was 103.8-105.3% for β-Actin and HER-2 respectively while CV value was around 1.2%. Our result showed a significant correlation with reported methods, which could potentially complement with FDA approved methods.

show abstract

Realtime quantification of HER2/neu gene amplification by polymerase chain reaction (PCR)

Cited by 2 publications

References 0 publications

Data provision of PIK3CA gene diversity and recombinant plasmids preparation for control DNA in developing the trastuzumab predictive response diagnostic kit

Data provision of PIK3CA gene diversity and recombinant plasmids preparation for control DNA in developing the trastuzumab predictive response diagnostic kit

Housekeeping gene as a source of calibrator candidate for HER-2 scoring in frozen tissue breast cancer study based on qPCR

Contact Info

Product

Resources

About