Comparison of the IHC, FISH, SISH and qPCR methods for the molecular diagnosis of breast cancer

Tvrdík, Daniel; Staněk, Libor; Skálová, Helena; Dundr, Pavel; Velenská, Z; Povýšil, C

doi:10.3892/mmr.2012.919

Cited by 17 publications

(12 citation statements)

References 9 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Furthermore, qPCR has been demonstrated to be a valuable tool for the evalu ation of HER2 gene amplification. These results are similar to those of a previous extensive and multicentric study focusing on the comparison of various ISH methods and qPCR in breast cancer, as well as being comparable with a previous study on breast carcinoma (10,26).…”

Section: Values -----------------------------------------------------supporting

confidence: 93%

Comparison of immunohistochemistry, four in situ hybridization methods and quantitative polymerase chain reaction for the molecular diagnosis of HER2 status in gastric cancer: A study of 55 cases

Staněk

Rozkoš

Laco

et al. 2014

Molecular Medicine Reports

View full text Add to dashboard Cite

In the current study, the sensitivity and specificity of methods of HER2 status detection were studied in 55 patients presenting with gastric/gastroesophageal junction carcinoma (30 intestinal and 25 diffuse), in small biopsy (endoscopy; n=33) and resection specimens (n=22). The primary objective of the present study was to compare various methods for the assessment of HER2 status, with regards to the sensitivity and specificity of each method, as well as their concordance. In all cases, the status of HER2 was determined using immunohistochemistry (IHC), fluorescence in situ hybridization (FISH), silver in situ hybridization (SISH), and quantitative polymerase chain reaction (qPCR). The concordance rate between IHC and ISH was 100% for IHC 0 and 3+. The concordance rate for IHC 1+ was 100% between IHC and SISH, and 92.9% between IHC and FISH. The concordance rate among different FISH methods was 100%, between FISH and SISH it was 96.2%, and between qPCR and ISH methods it was 88.5%. Thus, the results demonstrate that different in situ hybridization methods are comparable and that none were superior. Furthermore, the IHC and FISH methods were found to be comparable and the concordance rate was particularly good. qPCR analysis correlated well with the other methods and appears to be a possible alternative tool for detection of the HER2 status. However, the concordance rate of qPCR with other methods was identified to be lower in the diffuse carcinoma group of endoscopy biopsy specimens; therefore investigation of further cases is required.

show abstract

Section: Values -----------------------------------------------------supporting

confidence: 93%

Comparison of immunohistochemistry, four in situ hybridization methods and quantitative polymerase chain reaction for the molecular diagnosis of HER2 status in gastric cancer: A study of 55 cases

Staněk

Rozkoš

Laco

et al. 2014

Molecular Medicine Reports

View full text Add to dashboard Cite

show abstract

“…neoadjuvant therapy), which might influence the regulation and activity of the PI3K signalling pathway (Gonzalez-Angulo et al 2011). Third, while most other studies used IHC (Perren et al 1999; Depowski et al 2001; Lin et al 2003; Lee et al 2004; Engin et al 2006; Aleskandarany et al 2010; Gonzalez-Angulo et al 2011), we used qPCR to determine expression of PTEN and PIK3CA, which enables quantification relative to a household gene and is not biased by subjective factors such as the experience of the assessor (Tvrdik et al 2012; Mendoza et al 2013). …”

Section: Discussionmentioning

confidence: 99%

Expression of PIK3CA, PTEN mRNA and PIK3CA mutations in primary breast cancer: association with lymph node metastases

et al. 2013

View full text Add to dashboard Cite

PurposeHigh activity of the intracellular phosphatidylinositol-3 kinase (PI3K) pathway is common in breast cancer. Here, we explore differences in expression of important PI3K pathway regulators: the activator, phosphatidylinositol-3-kinase catalytic subunit alpha (PIK3CA), and the tumour suppressor, phosphatase and tensin homolog (PTEN), in breast carcinoma tissue and normal breast tissue. Furthermore, we examine whether expression of PIK3CA and PTEN mRNA and occurrence of PIK3CA mutations are associated with lymph node metastases in patients with primary breast cancer.MethodsPaired tissue samples of breast carcinoma and normal breast tissue were obtained from 175 breast cancer patients at the time of primary surgery, of these 105 patients were lymph node positive. Expression of PIK3CA and PTEN mRNA was quantified with Quantitative Real Time PCR. Somatic mutations in exon 9 and exon 20 of the PIK3CA gene were identified by genotyping.ResultsBoth PIK3CA and PTEN mRNA expression was significantly increased in breast carcinoma tissue compared to normal breast tissue (p = 2 × 10-11) and (p < 0.001), respectively. PIK3CA mutations were present in 68 out of 175 patients (39%), but were not associated with PIK3CA expression (p = 0.59). Expression of PIK3CA and PTEN mRNA, and PIK3CA mutations in breast carcinomas were not associated with presence of lymph node metastases.ConclusionsThe expression of PTEN and PIK3CA mRNA is increased in breast carcinoma tissue compared to normal breast tissue, and PIK3CA mutations are frequent in primary breast carcinoma, however these factors were not associated with lymph node metastases.Electronic supplementary materialThe online version of this article (doi:10.1186/2193-1801-2-464) contains supplementary material, which is available to authorized users.

show abstract

“…As few as 50 cells extracted from archival formalin-fixed paraffin-embedded (FFPE) tissues may be quantified using quantitative PCR (qPCR) [9] , [10] . The results from qPCR of HER2 measurements have been positively correlated with the results from IHC and FISH analysis [10] , [11] , [12] , [13] . Koudelakova et al compared qPCR to IHC and FISH data in breast cancer samples, and found that high sensitivity and specificity of the new method was achieved and the results obtained with the qPCR method and FISH/IHC agreed [14] .…”

Section: Introductionmentioning

confidence: 99%

Development of NIST standard reference material 2373: Genomic DNA standards for HER2 measurements

Almeida

Lund

et al. 2016

Biomolecular Detection and Quantification

View full text Add to dashboard Cite

NIST standard reference material (SRM) 2373 was developed to improve the measurements of the HER2 gene amplification in DNA samples. SRM 2373 consists of genomic DNA extracted from five breast cancer cell lines with different amounts of amplification of the HER2 gene. The five components are derived from the human cell lines SK-BR-3, MDA-MB-231, MDA-MB-361, MDA-MB-453, and BT-474. The certified values are the ratios of the HER2 gene copy numbers to the copy numbers of selected reference genes DCK, EIF5B, RPS27A, and PMM1. The ratios were measured using quantitative polymerase chain reaction and digital PCR, methods that gave similar ratios. The five components of SRM 2373 have certified HER2 amplification ratios that range from 1.3 to 17.7. The stability and homogeneity of the reference materials were shown by repeated measurements over a period of several years. SRM 2373 is a well characterized genomic DNA reference material that can be used to improve the confidence of the measurements of HER2 gene copy number.

show abstract

Comparison of the IHC, FISH, SISH and qPCR methods for the molecular diagnosis of breast cancer

Cited by 17 publications

References 9 publications

Comparison of immunohistochemistry, four in situ hybridization methods and quantitative polymerase chain reaction for the molecular diagnosis of HER2 status in gastric cancer: A study of 55 cases

Comparison of immunohistochemistry, four in situ hybridization methods and quantitative polymerase chain reaction for the molecular diagnosis of HER2 status in gastric cancer: A study of 55 cases

Expression of PIK3CA, PTEN mRNA and PIK3CA mutations in primary breast cancer: association with lymph node metastases

Development of NIST standard reference material 2373: Genomic DNA standards for HER2 measurements

Contact Info

Product

Resources

About