Chromogenic in situ hybridization

Last updated

Chromogenic in situ hybridization (CISH) is a cytogenetic technique that combines the chromogenic signal detection method of immunohistochemistry (IHC) techniques with in situ hybridization. [1] [2] It was developed around the year 2000 as an alternative to fluorescence in situ hybridization (FISH) for detection of HER-2/neu oncogene amplification. [1] CISH is similar to FISH in that they are both in situ hybridization techniques used to detect the presence or absence of specific regions of DNA. [1] However, CISH is much more practical in diagnostic laboratories because it uses bright-field microscopes rather than the more expensive and complicated fluorescence microscopes used in FISH. [1] [3]

Contents

Procedure

Procedure for performing chromogenic in situ hybridization CISH workflow.png
Procedure for performing chromogenic in situ hybridization

Probe design

Probe design for CISH is very similar to that for FISH with differences only in labelling and detection. FISH probes are generally labelled with a variety of different fluorescent tags and can only be detected under a fluorescence microscope, [4] whereas CISH probes are labelled with biotin or digoxigenin [5] and can be detected using a bright-field microscope after other treatment steps have been applied. [1]

CISH probes are approximately 20 nucleotides in length and are designed for DNA targets. They are complementary to the targeted sequence and bind to it after a denaturation and hybridization step. Only a few CISH probes are available commercially, so for most applications they have to be extracted, amplified, sequenced, labelled and mapped from bacterial artificial chromosomes (BACs). [6] BACs were developed during the Human Genome Project as it was necessary to isolate and amplify short fragments of human DNA for sequencing purposes. [7] Nowadays, BACs can be selected and positioned on the human genome using public databases such as the UCSC Genome Browser. [6] This ensures optimal complementarity and sequence specificity. DNA is extracted from the BAC clones and amplified using a polymerase-based technique, such as degenerate oligonucleotide primed (DOP)-PCR. [8] Next, the clones are sequenced and their position on the genome is verified. [9] Probe labelling can be carried out by using either random priming or nick translation to incorporate biotin or digoxigenin. [10]

Preparation of tissue, hybridization of probes, and detection

For CISH to work optimally, chromosomes must be in either interphase or metaphase. Tissue samples are securely attached to a surface, which is usually a glass slide, with paraffin. [11] The tissue samples must then be washed and heated several times to remove any paraffin before the hybridization step. After this, the sample has to undergo pepsin digestion to ensure the target is accessible. [11] As a final step, 10–20 μL of probe is added, the sample is covered with a coverslip which is sealed with rubber cement, and the slide is heated to 97 °C for 5–10 minutes to denature the DNA. [11] The slide is then placed in a 37 °C oven overnight so that the probe can hybridize. [11] [12] On the next day, the sample is washed and a blocker for nonspecific protein binding sites is applied. [11] If horseradish peroxidase (HRP) is going to be used, the sample must be incubated in hydrogen peroxide to suppress endogenous peroxidase activity. [11] If digoxigenin was used as a probe label, an anti-digoxigenin fluorescein primary antibody followed by a HRP-conjugated anti-fluorescein secondary antibody are then applied. [1] If biotin was used as a probe label, non-specific binding sites must first be blocked using bovine serum albumin (BSA). [11] Then, HRP-conjugated streptavidin is used for detection. [6] [11] HRP then converts diaminobenzidine (DAB) into an insoluble brown product, which can be detected in a bright-field microscope under 40- to 60-fold magnification. [11] [13] Staining lightly with hematoxylin can be used as a counterstain to make the product more visible. [5]

Comparison to other techniques

A) Direct FISH detection. Fluorescent labels are attached to a probe which will hybridize to a target DNA strand. B) Indirect FISH detection. Biotin, for example, is attached to a probe. Streptavidin linked to a fluorescent tag binds biotin with high specificity. C) Indirect CISH detection. Again, biotin is attached to a probe. Streptavidin linked to horseradish peroxidase binds biotin with high specificity. Horseradish peroxidase converts diaminobenzidine into a brown precipitate. FISH versus CISH Detection.png
A) Direct FISH detection. Fluorescent labels are attached to a probe which will hybridize to a target DNA strand. B) Indirect FISH detection. Biotin, for example, is attached to a probe. Streptavidin linked to a fluorescent tag binds biotin with high specificity. C) Indirect CISH detection. Again, biotin is attached to a probe. Streptavidin linked to horseradish peroxidase binds biotin with high specificity. Horseradish peroxidase converts diaminobenzidine into a brown precipitate.

Compared to FISH

FISH is considered to be the gold standard for the detection of chromosomal abnormalities because it is very sensitive and has high resolution. [3] [14] Other techniques that are developed to detect chromosomal abnormalities are usually compared to the sensitivity and specificity of FISH to see how they measure up. [3] [14] For example, compared to FISH, CISH has been shown to have a sensitivity of 97.5% and a specificity of 94% for detection of HER-2/neu gene amplification. [3] The concordance rate between FISH and CISH was 94.8%, showing CISH to be a comparable technique to FISH. [3] Most other sources agree and report an almost equal performance on gene amplification assays for FISH and CISH. [15] [16] However, sometimes CISH shows lower sensitivity for low level amplifications. [1]

CISH has some advantages over FISH in the reagents and equipment it uses. As noted above, CISH is much cheaper and is easier to use because it uses bright-field microscopes instead of fluorescence microscopes. [1] [3] In addition, the CISH reagents are more stable than the FISH reagents so it is possible to store the samples and examine the same sample multiple times. [3] [14] FISH reagents fade over time due to photobleaching so a sample can only be examined once. [3] [14] Apart from the expensive fluorescence microscope, FISH also requires a high-resolution digital camera to capture micrographs of the sample before the fluorescence fades. [14] Another advantage of using bright-field microscopy is that the tissue or cell sample as a whole can be visualized through CISH whereas cell morphology is difficult to assess using fluorescence microscopy in FISH. [3] [14]

CISH also differs from FISH in the probes that are used as well as in the overall method. There are many different types of FISH probes available, such as repeat probes, probes that detect specific genes or telomeres, and probes that detect chromosomal abnormalities. [14] In contrast, there is a limited variety of commercially available CISH probes, including probes that bind the centromere of chromosomes 3, 7, 8, 9, 10, 11, 17, 18, X, and Y as well as gene-specific probes for cancer-related genes, such as HER-2, EGFR, MYC, and TOP2A. [14] Despite the limited variety of available CISH probes, they are generally more cost-effective than FISH probes. [14] With regard to the overall method, FISH can be performed using direct labelling—fluorochromes are attached to the probes—or indirect labelling—the probes are labelled with biotin or digoxigenin which are then detected using fluorescently-labelled streptavidin or antibodies, respectively. [14] CISH is performed using indirect labelling in which antibodies or streptavidin are conjugated to enzymes such as HRP or alkaline phosphatase (AP). [14]

Compared to IHC

CISH and IHC are similar in that both are used for the same purpose (mainly to detect HER-2/neu amplification) and they both use enzyme reactions (HRP/AP) to measure amplification. [13] CISH and IHC are different in that IHC measures protein expression whereas CISH measures DNA amplification. [13] This difference is particularly useful for HER-2/neu because it has been found that gene amplification is of higher prognostic value than protein expression. [17]

A disadvantage of IHC is that it is not possible to identify false-negative and false-positive results. [17] In CISH, if there is no signal for the reference probe, the assay has failed.

For low and high protein overexpression/gene amplification, CISH and IHC show a concordance of over 86% and over 89%, respectively. [18] It has been shown that monoclonal antibodies are better than polyclonal antibodies for detection in both IHC and CISH as they bind more specifically, which leads to a higher concordance rate. [18]

For medium protein overexpression/gene amplification concordance varies, but is higher when monoclonal antibodies are used than when polyclonal antibodies are used. [18] The variable concordance is due to the fact that gene amplification does not strictly correlate with protein expression and that tumor heterogeneity can make it difficult to detect protein overexpression in a tissue. [18]

Medical applications

CISH is frequently applied to assess gene amplification, such as HER-2/neu status in breast cancer samples. [2] [19] HER-2/neu amplification is associated with higher mortality, higher recurrence rate, and poor prognosis in breast cancer. [20] The monoclonal antibody trastuzumab is a receptor blocker that has been proven to be clinically very effective in HER-2/neu-overexpressing tumors. [21] Therefore, it is crucial to determine receptor status before starting cancer treatment. [21]

CISH is also used for detection of chromosomal rearrangements and fusions, such as the fusion of ALK tyrosine kinase domain with the promoter and 5’ region of EML4 in lung cancer. ALK-positive tumors are a clinically relevant subgroup as they can be very effectively treated with the ALK inhibitor crizotinib. [22] [23]

Apart from cancers, CISH has also been shown to be useful in detecting human papillomavirus infections. [24]

Variations

Silver-enhanced in situ hybridization (SISH)

SISH uses a similar method as CISH, but a silver precipitate is the end product rather than a brown product. [25] In this method, a probe tagged with dinitrophenol (DNP) binds to the target sequence. [25] A primary anti-DNP antibody is then added followed by a secondary antibody conjugated to HRP. [25] Silver acetate, hydroquinone, and hydrogen peroxide are subsequently added to the secondary antibody and HRP catalyzes the polymerization of silver in the presence of hydrogen peroxide. [25] Silver metal is consequently deposited into the nuclei of the cell. [25] Amplification of HER-2/neu is seen as black dots. [25]

DuoCISH

DuoCISH is a variation of CISH that addresses the need for two different probes on the same slide. [26] It is a well-established technique for HER-2/neu amplification detection even though it is sometimes reported to be less effective than FISH. [27] In this technique, one probe binds the reference which, in the case of HER-2/neu amplification detection, is CEN17 (the centromere of chromosome 17) while the other probe binds the target sequence which is HER-2/neu. [26] [27] DuoCISH combines both FISH and CISH in that it converts signals from FISH probes to chromogenic substrates. [26] [27] It works on the principle that blue cyanine dye is a substrate for HRP and red dye is a substrate for AP. For example, green fluorescein isothiocyanate (FITC) signals are converted to blue chromogenic precipitates through an anti-FITC antibody conjugated to HRP, and Texas Red signals are converted to red chromogenic precipitates through an anti-Texas Red antibody conjugated to AP. [26] [27] An advantage of DuoCISH is that it is possible to distinguish between chromosomal aneuploidy and gene amplification as the reference gene will also be amplified in aneuploidy but not in gene amplification detection. [26] [27]

Related Research Articles

<span class="mw-page-title-main">Trastuzumab</span> Pharmaceutical drug

Trastuzumab, sold under the brand name Herceptin among others, is a monoclonal antibody used to treat breast cancer and stomach cancer. It is specifically used for cancer that is HER2 receptor positive. It may be used by itself or together with other chemotherapy medication. Trastuzumab is given by slow injection into a vein and injection just under the skin.

Comparative genomic hybridization(CGH) is a molecular cytogenetic method for analysing copy number variations (CNVs) relative to ploidy level in the DNA of a test sample compared to a reference sample, without the need for culturing cells. The aim of this technique is to quickly and efficiently compare two genomic DNA samples arising from two sources, which are most often closely related, because it is suspected that they contain differences in terms of either gains or losses of either whole chromosomes or subchromosomal regions (a portion of a whole chromosome). This technique was originally developed for the evaluation of the differences between the chromosomal complements of solid tumor and normal tissue, and has an improved resolution of 5–10 megabases compared to the more traditional cytogenetic analysis techniques of giemsa banding and fluorescence in situ hybridization (FISH) which are limited by the resolution of the microscope utilized.

<span class="mw-page-title-main">Immunohistochemistry</span> Common application of immunostaining

Immunohistochemistry (IHC) is the most common application of immunostaining. It involves the process of selectively identifying antigens (proteins) in cells of a tissue section by exploiting the principle of antibodies binding specifically to antigens in biological tissues. IHC takes its name from the roots "immuno", in reference to antibodies used in the procedure, and "histo", meaning tissue. Albert Coons conceptualized and first implemented the procedure in 1941.

In molecular biology, a hybridization probe(HP) is a fragment of DNA or RNA of usually 15–10000 nucleotide long which can be radioactively or fluorescently labeled. HP can be used to detect the presence of nucleotide sequences in analyzed RNA or DNA that are complementary to the sequence in the probe. The labeled probe is first denatured (by heating or under alkaline conditions such as exposure to sodium hydroxide) into single stranded DNA (ssDNA) and then hybridized to the target ssDNA (Southern blotting) or RNA (northern blotting) immobilized on a membrane or in situ.

Fluorescence <i>in situ</i> hybridization Genetic testing technique

Fluorescence in situ hybridization (FISH) is a molecular cytogenetic technique that uses fluorescent probes that bind to only particular parts of a nucleic acid sequence with a high degree of sequence complementarity. It was developed by biomedical researchers in the early 1980s to detect and localize the presence or absence of specific DNA sequences on chromosomes. Fluorescence microscopy can be used to find out where the fluorescent probe is bound to the chromosomes. FISH is often used for finding specific features in DNA for use in genetic counseling, medicine, and species identification. FISH can also be used to detect and localize specific RNA targets in cells, circulating tumor cells, and tissue samples. In this context, it can help define the spatial-temporal patterns of gene expression within cells and tissues.

<i>In situ</i> hybridization

In situ hybridization (ISH) is a type of hybridization that uses a labeled complementary DNA, RNA or modified nucleic acids strand to localize a specific DNA or RNA sequence in a portion or section of tissue or if the tissue is small enough, in the entire tissue, in cells, and in circulating tumor cells (CTCs). This is distinct from immunohistochemistry, which usually localizes proteins in tissue sections.

<span class="mw-page-title-main">Polysomy</span> Abnormal multiples of one or more chromosomes

Polysomy is a condition found in many species, including fungi, plants, insects, and mammals, in which an organism has at least one more chromosome than normal, i.e., there may be three or more copies of the chromosome rather than the expected two copies. Most eukaryotic species are diploid, meaning they have two sets of chromosomes, whereas prokaryotes are haploid, containing a single chromosome in each cell. Aneuploids possess chromosome numbers that are not exact multiples of the haploid number and polysomy is a type of aneuploidy. A karyotype is the set of chromosomes in an organism and the suffix -somy is used to name aneuploid karyotypes. This is not to be confused with the suffix -ploidy, referring to the number of complete sets of chromosomes.

<span class="mw-page-title-main">HER2</span> Mammalian protein found in humans

Receptor tyrosine-protein kinase erbB-2 is a protein that normally resides in the membranes of cells and is encoded by the ERBB2 gene. ERBB is abbreviated from erythroblastic oncogene B, a gene originally isolated from the avian genome. The human protein is also frequently referred to as HER2 or CD340.

<span class="mw-page-title-main">Molecular cytogenetics</span>

Molecular cytogenetics combines two disciplines, molecular biology and cytogenetics, and involves the analysis of chromosome structure to help distinguish normal and cancer-causing cells. Human cytogenetics began in 1956 when it was discovered that normal human cells contain 46 chromosomes. However, the first microscopic observations of chromosomes were reported by Arnold, Flemming, and Hansemann in the late 1800s. Their work was ignored for decades until the actual chromosome number in humans was discovered as 46. In 1879, Arnold examined sarcoma and carcinoma cells having very large nuclei. Today, the study of molecular cytogenetics can be useful in diagnosing and treating various malignancies such as hematological malignancies, brain tumors, and other precursors of cancer. The field is overall focused on studying the evolution of chromosomes, more specifically the number, structure, function, and origin of chromosome abnormalities. It includes a series of techniques referred to as fluorescence in situ hybridization, or FISH, in which DNA probes are labeled with different colored fluorescent tags to visualize one or more specific regions of the genome. Introduced in the 1980s, FISH uses probes with complementary base sequences to locate the presence or absence of the specific DNA regions. FISH can either be performed as a direct approach to metaphase chromosomes or interphase nuclei. Alternatively, an indirect approach can be taken in which the entire genome can be assessed for copy number changes using virtual karyotyping. Virtual karyotypes are generated from arrays made of thousands to millions of probes, and computational tools are used to recreate the genome in silico.

Multiplex ligation-dependent probe amplification (MLPA) is a variation of the multiplex polymerase chain reaction that permits amplification of multiple targets with only a single primer pair. It detects copy number changes at the molecular level, and software programs are used for analysis. Identification of deletions or duplications can indicate pathogenic mutations, thus MLPA is an important diagnostic tool used in clinical pathology laboratories worldwide.

Digital polymerase chain reaction is a biotechnological refinement of conventional polymerase chain reaction methods that can be used to directly quantify and clonally amplify nucleic acids strands including DNA, cDNA, or RNA. The key difference between dPCR and traditional PCR lies in the method of measuring nucleic acids amounts, with the former being a more precise method than PCR, though also more prone to error in the hands of inexperienced users. A "digital" measurement quantitatively and discretely measures a certain variable, whereas an “analog” measurement extrapolates certain measurements based on measured patterns. PCR carries out one reaction per single sample. dPCR also carries out a single reaction within a sample, however the sample is separated into a large number of partitions and the reaction is carried out in each partition individually. This separation allows a more reliable collection and sensitive measurement of nucleic acid amounts. The method has been demonstrated as useful for studying variations in gene sequences — such as copy number variants and point mutations — and it is routinely used for clonal amplification of samples for next-generation sequencing.

<span class="mw-page-title-main">ZNF217</span> Protein-coding gene in the species Homo sapiens

Zinc finger protein 217, also known as ZNF217, is a protein which in humans is encoded by the ZNF217 gene.

A Riboprobe, abbreviation of RNA probe, is a segment of labelled RNA that can be used to detect a target mRNA or DNA during in situ hybridization. RNA probes can be produced by in vitro transcription of cloned DNA inserted in a suitable plasmid downstream of a viral promoter. Some bacterial viruses code for their own RNA polymerases, which are highly specific for the viral promoters. Using these enzymes, labeled NTPs, and inserts inserted in both forward and reverse orientations, both sense and antisense riboprobes can be generated from a cloned gene.

Virtual karyotype is the digital information reflecting a karyotype, resulting from the analysis of short sequences of DNA from specific loci all over the genome, which are isolated and enumerated. It detects genomic copy number variations at a higher resolution for level than conventional karyotyping or chromosome-based comparative genomic hybridization (CGH). The main methods used for creating virtual karyotypes are array-comparative genomic hybridization and SNP arrays.

Molecular Inversion Probe (MIP) belongs to the class of Capture by Circularization molecular techniques for performing genomic partitioning, a process through which one captures and enriches specific regions of the genome. Probes used in this technique are single stranded DNA molecules and, similar to other genomic partitioning techniques, contain sequences that are complementary to the target in the genome; these probes hybridize to and capture the genomic target. MIP stands unique from other genomic partitioning strategies in that MIP probes share the common design of two genomic target complementary segments separated by a linker region. With this design, when the probe hybridizes to the target, it undergoes an inversion in configuration and circularizes. Specifically, the two target complementary regions at the 5’ and 3’ ends of the probe become adjacent to one another while the internal linker region forms a free hanging loop. The technology has been used extensively in the HapMap project for large-scale SNP genotyping as well as for studying gene copy alterations and characteristics of specific genomic loci to identify biomarkers for different diseases such as cancer. Key strengths of the MIP technology include its high specificity to the target and its scalability for high-throughput, multiplexed analyses where tens of thousands of genomic loci are assayed simultaneously.

<span class="mw-page-title-main">Proximity ligation assay</span>

Proximity ligation assay is a technology that extends the capabilities of traditional immunoassays to include direct detection of proteins, protein interactions, extracellular vesicles and post translational modifications with high specificity and sensitivity. Protein targets can be readily detected and localized with single molecule resolution and objectively quantified in unmodified cells and tissues. Utilizing only a few cells, sub-cellular events, even transient or weak interactions, are revealed in situ and sub-populations of cells can be differentiated. Within hours, results from conventional co-immunoprecipitation and co-localization techniques can be confirmed.

<span class="mw-page-title-main">Molecular diagnostics</span> Collection of techniques used to analyze biological markers in the genome and proteome

Molecular diagnostics is a collection of techniques used to analyze biological markers in the genome and proteome, and how their cells express their genes as proteins, applying molecular biology to medical testing. In medicine the technique is used to diagnose and monitor disease, detect risk, and decide which therapies will work best for individual patients, and in agricultural biosecurity similarly to monitor crop- and livestock disease, estimate risk, and decide what quarantine measures must be taken.

<span class="mw-page-title-main">Neuronal lineage marker</span> Endogenous tag expressed in different cells along neurogenesis and differentiated cells

A neuronal lineage marker is an endogenous tag that is expressed in different cells along neurogenesis and differentiated cells such as neurons. It allows detection and identification of cells by using different techniques. A neuronal lineage marker can be either DNA, mRNA or RNA expressed in a cell of interest. It can also be a protein tag, as a partial protein, a protein or an epitope that discriminates between different cell types or different states of a common cell. An ideal marker is specific to a given cell type in normal conditions and/or during injury. Cell markers are very valuable tools for examining the function of cells in normal conditions as well as during disease. The discovery of various proteins specific to certain cells led to the production of cell-type-specific antibodies that have been used to identify cells.

Mammary analogue secretory carcinoma (MASC), also termed MASCSG, is a salivary gland neoplasm. It is a secretory carcinoma which shares the microscopic pathologic features with other types of secretory carcinomas including mammary secretory carcinoma, secretory carcinoma of the skin, and salivary gland–type carcinoma of the thyroid. MASCSG was first described by Skálová et al. in 2010. The authors of this report found a chromosome translocation in certain salivary gland tumors, i.e. a (12;15)(p13;q25) fusion gene mutation. The other secretory carcinoma types carry this fusion gene.

<span class="mw-page-title-main">Spatial transcriptomics</span> Range of methods designed for assigning cell types

Spatial transcriptomics is a method for assigning cell types to their locations in the histological sections and can also be used to determine subcellular localization of mRNA molecules. First described in 2016 by Ståhl et al., it has since undergone a variety of improvements and modifications.

References

  1. 1 2 3 4 5 6 7 8 Tanner, M; Gancberg, D; Di Leo, A; Larsimont, D; Rouas, G; Piccart, M. J.; Isola, J (2000). "Chromogenic in situ hybridization: A practical alternative for fluorescence in situ hybridization to detect HER-2/neu oncogene amplification in archival breast cancer samples". The American Journal of Pathology. 157 (5): 1467–72. doi:10.1016/S0002-9440(10)64785-2. PMC   1885742 . PMID   11073807.
  2. 1 2 Madrid, M. A.; Lo, R. W. (2004). "Chromogenic in situ hybridization (FISH): A novel alternative in screening archival breast cancer tissue samples for HER-2/neu status". Breast Cancer Research. 6 (5): R593–600. doi: 10.1186/bcr915 . PMC   549176 . PMID   15318940.
  3. 1 2 3 4 5 6 7 8 9 Sáez, A; Andreu, F. J.; Seguí, M. A.; Baré, M. L.; Fernández, S; Dinarés, C; Rey, M (2006). "HER-2 gene amplification by chromogenic in situ hybridisation (CISH) compared with fluorescence in situ hybridisation (FISH) in breast cancer-A study of two hundred cases". The Breast. 15 (4): 519–27. doi:10.1016/j.breast.2005.09.008. PMID   16290155.
  4. ↑ Garimberti, E; Tosi, S (2010). "Fluorescence in situ Hybridization (FISH), Basic Principles and Methodology". Fluorescence in situ Hybridization (FISH). Methods in Molecular Biology. Vol. 659. pp. 3–20. doi:10.1007/978-1-60761-789-1_1. ISBN   978-1-60761-788-4. PMID   20809300.
  5. 1 2 Lambros, M. B.; Simpson, P. T.; Jones, C; Natrajan, R; Westbury, C; Steele, D; Savage, K; MacKay, A; Schmitt, F. C.; Ashworth, A; Reis-Filho, J. S. (2006). "Unlocking pathology archives for molecular genetic studies: A reliable method to generate probes for chromogenic and fluorescent in situ hybridization". Laboratory Investigation. 86 (4): 398–408. doi: 10.1038/labinvest.3700390 . PMID   16446704.
  6. 1 2 3 Summersgill, B. M.; Shipley, J. M. (2010). "Fluorescence in Situ Hybridization Analysis of Formalin Fixed Paraffin Embedded Tissues, Including Tissue Microarrays". Fluorescence in situ Hybridization (FISH). Methods in Molecular Biology. Vol. 659. pp. 51–70. doi:10.1007/978-1-60761-789-1_4. ISBN   978-1-60761-788-4. PMID   20809303.
  7. ↑ Shizuya, H; Kouros-Mehr, H (2001). "The development and applications of the bacterial artificial chromosome cloning system". The Keio Journal of Medicine. 50 (1): 26–30. doi: 10.2302/kjm.50.26 . PMID   11296661.
  8. ↑ Darouich, S; Popovici, C; Missirian, C; Moncla, A (2012). "Use of DOP-PCR for amplification and labeling of BAC DNA for FISH". Biotechnic & Histochemistry. 87 (2): 117–21. doi:10.3109/10520295.2011.559175. PMID   21314248. S2CID   1326145.
  9. ↑ De Braekeleer, E; Douet-Guilbert, N; Basinko, A; Morel, F; Le Bris, M. J.; Férec, C; De Braekeleer, M (2011). "Using bacterial artificial chromosomes in leukemia research: The experience at the university cytogenetics laboratory in Brest, France". Journal of Biomedicine and Biotechnology. 2011: 1–7. doi: 10.1155/2011/329471 . PMC   3025366 . PMID   21274439.
  10. ↑ Cold Spring Harbor Laboratory Press. (2012) "Labeling of DNA Probes by Nick Translation". Molecular Cloning: A Laboratory Manual.
  11. 1 2 3 4 5 6 7 8 9 Park, K; Kim, J; Lim, S; Han, S; Lee, J. Y. (2003). "Comparing fluorescence in situ hybridization and chromogenic in situ hybridization methods to determine the HER2/neu status in primary breast carcinoma using tissue microarray". Modern Pathology. 16 (9): 937–43. doi: 10.1097/01.MP.0000086487.78558.7D . PMID   13679458.
  12. ↑ Schildhaus, H. U.; Deml, K. F.; Schmitz, K; Meiboom, M; Binot, E; Hauke, S; Merkelbach-Bruse, S; Büttner, R (2013). "Chromogenic in situ hybridization is a reliable assay for detection of ALK rearrangements in adenocarcinomas of the lung". Modern Pathology. 26 (11): 1468–77. doi: 10.1038/modpathol.2013.95 . PMID   23743932.
  13. 1 2 3 Gupta, D; Middleton, L. P.; Whitaker, M. J.; Abrams, J (2003). "Comparison of fluorescence and chromogenic in situ hybridization for detection of HER-2/neu oncogene in breast cancer". American Journal of Clinical Pathology. 119 (3): 381–7. doi: 10.1309/P40P2EAD42PUKDMG . PMID   12645340.
  14. 1 2 3 4 5 6 7 8 9 10 11 Lambros, M. B.; Natrajan, R; Reis-Filho, J. S. (2007). "Chromogenic and fluorescent in situ hybridization in breast cancer". Human Pathology. 38 (8): 1105–22. doi:10.1016/j.humpath.2007.04.011. PMID   17640550.
  15. ↑ Zhao, J; Wu, R; Au, A; Marquez, A; Yu, Y; Shi, Z (2002). "Determination of HER2 gene amplification by chromogenic in situ hybridization (CISH) in archival breast carcinoma". Modern Pathology. 15 (6): 657–65. doi: 10.1038/modpathol.3880582 . PMID   12065780. S2CID   19329525.
  16. ↑ Shia, J; Klimstra, D. S.; Li, A. R.; Qin, J; Saltz, L; Teruya-Feldstein, J; Akram, M; Chung, K. Y.; Yao, D; Paty, P. B.; Gerald, W; Chen, B (2005). "Epidermal growth factor receptor expression and gene amplification in colorectal carcinoma: An immunohistochemical and chromogenic in situ hybridization study". Modern Pathology. 18 (10): 1350–6. doi: 10.1038/modpathol.3800417 . PMID   15832190.
  17. 1 2 Todorović-Raković, N; Jovanović, D; Nesković-Konstantinović, Z; Nikolić-Vukosavljević, D (2005). "Comparison between immunohistochemistry and chromogenic in situ hybridization in assessing HER-2 status in breast cancer". Pathology International. 55 (6): 318–23. doi:10.1111/j.1440-1827.2005.01831.x. PMID   15943788. S2CID   24153338.
  18. 1 2 3 4 Rosa, F. E.; Santos, R. M.; Rogatto, S. R.; Domingues, M. A. (2013). "Chromogenic in situ hybridization compared with other approaches to evaluate HER2/neu status in breast carcinomas". Brazilian Journal of Medical and Biological Research. 46 (3): 207–16. doi:10.1590/1414-431x20132483. PMC   3854374 . PMID   23558859.
  19. ↑ Kumamoto, H; Sasano, H; Taniguchi, T; Suzuki, T; Moriya, T; Ichinohasama, R (2001). "Chromogenic in situ hybridization analysis of HER-2/neu status in breast carcinoma: Application in screening of patients for trastuzumab (Herceptin) therapy". Pathology International. 51 (8): 579–84. doi:10.1046/j.1440-1827.2001.01255.x. PMID   11564211. S2CID   21849968.
  20. ↑ Mitri, Z; Constantine, T; O'Regan, R (2012). "The HER2 Receptor in Breast Cancer: Pathophysiology, Clinical Use, and New Advances in Therapy". Chemotherapy Research and Practice. 2012: 1–7. doi: 10.1155/2012/743193 . PMC   3539433 . PMID   23320171.
  21. 1 2 Swain, S. M.; Kim, S. B.; Cortés, J; Ro, J; Semiglazov, V; Campone, M; Ciruelos, E; Ferrero, J. M.; Schneeweiss, A; Knott, A; Clark, E; Ross, G; Benyunes, M. C.; Baselga, J (2013). "Pertuzumab, trastuzumab, and docetaxel for HER2-positive metastatic breast cancer (CLEOPATRA study): Overall survival results from a randomised, double-blind, placebo-controlled, phase 3 study". The Lancet Oncology. 14 (6): 461–71. doi:10.1016/S1470-2045(13)70130-X. PMC   4076842 . PMID   23602601.
  22. ↑ Kwak, E. L.; Bang, Y. J.; Camidge, D. R.; Shaw, A. T.; Solomon, B; Maki, R. G.; Ou, S. H.; Dezube, B. J.; Jänne, P. A.; Costa, D. B.; Varella-Garcia, M; Kim, W. H.; Lynch, T. J.; Fidias, P; Stubbs, H; Engelman, J. A.; Sequist, L. V.; Tan, W; Gandhi, L; Mino-Kenudson, M; Wei, G. C.; Shreeve, S. M.; Ratain, M. J.; Settleman, J; Christensen, J. G.; Haber, D. A.; Wilner, K; Salgia, R; Shapiro, G. I.; et al. (2010). "Anaplastic lymphoma kinase inhibition in non-small-cell lung cancer". New England Journal of Medicine. 363 (18): 1693–703. doi:10.1056/NEJMoa1006448. PMC   3014291 . PMID   20979469.
  23. ↑ Wagner, F; Streubel, A; Roth, A; Stephan-Falkenau, S; Mairinger, T (2014). "Chromogenic in situ hybridisation (CISH) is a powerful method to detect ALK-positive non-small cell lung carcinomas". Journal of Clinical Pathology. 67 (5): 403–7. doi:10.1136/jclinpath-2013-201974. PMID   24293609. S2CID   25718718.
  24. ↑ Zappacosta, R; Colasante, A; Viola, P; d'Antuono, T; Lattanzio, G; Capanna, S; Gatta, D. M.; Rosini, S (2013). "Chromogenic in situ hybridization and p16/Ki67 dual staining on formalin-fixed paraffin-embedded cervical specimens: Correlation with HPV-DNA test, E6/E7 mRNA test, and potential clinical applications". BioMed Research International. 2013: 1–11. doi: 10.1155/2013/453606 . PMC   3858005 . PMID   24369532.
  25. 1 2 3 4 5 6 Shousha, S; Peston, D; Amo-Takyi, B; Morgan, M; Jasani, B (2009). "Evaluation of automated silver-enhanced in situ hybridization (SISH) for detection of HER2 gene amplification in breast carcinoma excision and core biopsy specimens". Histopathology. 54 (2): 248–53. doi:10.1111/j.1365-2559.2008.03185.x. PMID   19207950. S2CID   31938310.
  26. 1 2 3 4 5 Henriksen, U., Müller, S., and Schønau, A. (2009) Chapter 14: "Dual color CISH and FISH to CISH conversion", pp. 97–101 in G.L. Kumar and L. Rudbeck (Eds.), Immunohistochemical (IHC) Staining Methods Fifth Edition. Carpinteria, CA: Dako North America
  27. 1 2 3 4 5 Mollerup, J; Henriksen, U; Müller, S; Schønau, A (2012). "Dual color chromogenic in situ hybridization for determination of HER2 status in breast cancer: A large comparative study to current state of the art fluorescence in situ hybridization". BMC Clinical Pathology. 12: 3. doi: 10.1186/1472-6890-12-3 . PMC   3305592 . PMID   22333181.
This page is based on this Wikipedia article
Text is available under the CC BY-SA 4.0 license; additional terms may apply.
Images, videos and audio are available under their respective licenses.