{"id":649,"date":"2018-12-10T16:57:37","date_gmt":"2018-12-10T16:57:37","guid":{"rendered":"https:\/\/wp.uthscsa.edu\/pathology\/?page_id=649"},"modified":"2018-12-10T16:57:37","modified_gmt":"2018-12-10T16:57:37","slug":"fish-microdeletions","status":"publish","type":"page","link":"https:\/\/lsom.uthscsa.edu\/pathology\/reference-labs\/clinical-molecular-cytogenetics\/fish-microdeletions\/","title":{"rendered":"FISH \u2013 Microdeletions"},"content":{"rendered":"

[vc_row][vc_column width=”1\/2″][vc_column_text]<\/p>\n

DiGeorge\/VCFS\/22q11\u00a0Microdeletion<\/h3>\n

Test Overview<\/p>\n

CPT Code(s)<\/strong><\/p>\n

Peripheral Blood: 88230-52, 88271, 88273, 88291<\/p>\n

Amniotic Fluid: 88235-52, 88271, 88273, 88291<\/p>\n

Methodology<\/strong><\/p>\n

Culture\/Hybridization\/Microscopy\/Interpretation<\/p>\n

Specimen Requirements<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n
<\/td>\nPeripheral Blood<\/strong><\/td>\nAmniotic Fluid<\/strong><\/td>\n<\/tr>\n
Container<\/td>\nSodium Heparin (green top tube)<\/td>\nTwo 15 ml sterile leak-proof conical tubes<\/td>\n<\/tr>\n
Optimal Quantity<\/td>\n2-3 ml<\/td>\n20-25 ml<\/td>\n<\/tr>\n
Minimum Quantity<\/td>\n1-2 ml<\/td>\n10 ml<\/td>\n<\/tr>\n
Storage<\/td>\nRoom Temperature<\/td>\nRoom Temperature<\/td>\n<\/tr>\n
Stability at Room Temperature<\/td>\nRoom Temperature for 8 hours, then refrigerate<\/td>\nRoom Temperature for 8 hours, then refrigerate<\/td>\n<\/tr>\n
Transportation<\/td>\nAvoid freezing or heating over 35o<\/sup>C<\/td>\nAvoid freezing or heating over 35o<\/sup>C<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

Turnaround Time<\/strong><\/p>\n

Peripheral Blood: Final report in 5 days for 90% of cases<\/p>\n

Amniotic Fluid: Final report in 7-10 days for 90% of cases<\/p>\n

Test Details<\/p>\n

Clinical Significance<\/strong><\/p>\n

Fluorescence in situ hybridization (FISH) is a sensitive method to detect smaller genomic changes associated with various\u00a0micordeletion\u00a0syndromes.\u00a0\u00a0There are several advantages with FISH technology over routine chromosome analysis and such advantages include the ability of FISH technology to detect genomic abnormalities in non-viable and non-dividing tissues, rapid turnaround time, and increased resolution.\u00a0\u00a0However, FISH technology is complementary to routine chromosome analysis and cannot substitute routine chromosome analysis and FISH diagnostic testing is not recommended as an isolated test without chromosome analysis for the diagnosis of these\u00a0microdeletion\u00a0syndromes.<\/p>\n

Clinical Background<\/strong><\/p>\n

The 22q11\u00a0microdeletion\u00a0is associated with\u00a0DiGeorge\u00a0or\u00a0velocardiofacial\u00a0(VCFS) syndrome which is an autosomal dominant condition with variable expressivity resulting in variable phenotype.\u00a0\u00a0Patients with this deletion show characteristic features such as craniofacial anomalies, mental retardation, and congenital heart defect.\u00a0\u00a0Among the heart defects, tetralogy of\u00a0Fallot\u00a0is the most common.\u00a0\u00a0Several studies have recommended that any child with a congenital heart defect should be tested for 22q11\u00a0microdeletion\u00a0since this deletion is very common in about 20% of patients with\u00a0syndromic\u00a0heart defects.\u00a0\u00a0The 22q11\u00a0microdeletion\u00a0is also referred to as CATCH22 for cardiac defects, abnormal\u00a0facies,\u00a0thymic\u00a0hypoplasia, cleft palate and\u00a0hypocalcemia.\u00a0\u00a0Since the most common deletion encompasses ~3 Mb, often these deletions are not detected on routine chromosome analysis.\u00a0\u00a0FISH is the ideal choice for detection of this deletion due to its increased sensitivity.<\/p>\n

Methods<\/strong><\/p>\n

The sample is usually cultured until sufficient number of mitotic cells is observed (48-72 hours for peripheral blood and 7-10 days for amniotic fluid) and after harvesting the cells are hybridized with the probes.\u00a0\u00a0After hybridization and post-hybridization washes, the slides are analyzed under a fluorescence microscope equipped with\u00a0epifluorescence\u00a0and appropriate filter sets.\u00a0\u00a0Analysis and scoring is carried out by two certified technologists in a blinded fashion with each scoring a minimum of 5 metaphases and 50 interphase nuclei.\u00a0\u00a0Results are reviewed by both the laboratory manager and the director.<\/p>\n

22q11 deletion (DiGeorge\u00a0\/ VCFS syndrome)\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0HIRA \/ TUPLE1\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0Cytocell\u00a0(cat #LPU004)<\/p>\n

Interpretation of Results<\/strong><\/p>\n

The morphologic interpretation and correlation of results on all cases is performed by a board-certified doctoral level scientist (laboratory director).\u00a0The final report has a narrative description of results.\u00a0\u00a0If abnormalities are present, they are explained in a paragraph which helps to clarify and correlate chromosomal findings with phenotype.\u00a0References are included in the report to help the referring physician with interpretation, which include books or journals that contain appropriate information. Recommendations are made as to any additional testing, if necessary.<\/p>\n

<\/h3>\n

Prader-Willi Syndrome<\/h3>\n

Test Overview<\/p>\n

CPT Code(s)<\/strong><\/p>\n

Peripheral Blood: 88230-52, 88271, 88273, 88291<\/p>\n

Amniotic Fluid: 88235-52, 88271, 88273, 88291<\/p>\n

Methodology<\/strong><\/p>\n

Culture\/Hybridization\/Microscopy\/Interpretation<\/p>\n

Specimen Requirements<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n
<\/td>\nPeripheral Blood<\/strong><\/td>\nAmniotic Fluid<\/strong><\/td>\n<\/tr>\n
Container<\/td>\nSodium Heparin (green top tube)<\/td>\nTwo 15 ml sterile leak-proof conical tubes<\/td>\n<\/tr>\n
Optimal Quantity<\/td>\n2-3 ml<\/td>\n20-25 ml<\/td>\n<\/tr>\n
Minimum Quantity<\/td>\n1-2 ml<\/td>\n10 ml<\/td>\n<\/tr>\n
Storage<\/td>\nRoom Temperature<\/td>\nRoom Temperature<\/td>\n<\/tr>\n
Stability at Room Temperature<\/td>\nRoom Temperature for 8 hours, then refrigerate<\/td>\nRoom Temperature for 8 hours, then refrigerate<\/td>\n<\/tr>\n
Transportation<\/td>\nAvoid freezing or heating over 35o<\/sup>C<\/td>\nAvoid freezing or heating over 35o<\/sup>C<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

Turnaround Time<\/strong><\/p>\n

Peripheral Blood: Final report in 5 days for 90% of cases<\/p>\n

Amniotic Fluid: Final report in 7-10 days for 90% of cases<\/p>\n

Test Details<\/p>\n

Clinical Significance<\/strong><\/p>\n

Fluorescence in situ hybridization (FISH) is a sensitive method to detect smaller genomic changes associated with various\u00a0micordeletion\u00a0syndromes.\u00a0\u00a0There are several advantages with FISH technology over routine chromosome analysis and such advantages include the ability of FISH technology to detect genomic abnormalities in non-viable and non-dividing tissues, rapid turnaround time, and increased resolution.\u00a0\u00a0However, FISH technology is complementary to routine chromosome analysis and cannot substitute routine chromosome analysis and FISH diagnostic testing is not recommended as an isolated test without chromosome analysis for the diagnosis of these\u00a0microdeletion\u00a0syndromes.<\/p>\n

Clinical Background<\/strong><\/p>\n

Prader-Willi syndrome, caused by\u00a0microdeletions\/loss of paternally derived gene expression, is a\u00a0panethnic\u00a0developmental disorder affecting approximately 1 in 10,000 to 1 in 15,000 live births. Patients with this syndrome show characteristic features including unusual facial appearance, short stature, severe mental retardation, spasticity and seizures.\u00a0\u00a0Chromosome deletion 15q11q13 during male meiosis results in\u00a0Prader-Willi syndrome and such deletions account for a large number of patients while a small number of patients have mutations within this gene that cannot be detected by either routine chromosome analysis or FISH.\u00a0\u00a0\u00a0Since the most common deletion encompasses ~3 Mb, often these deletions are not detected on routine chromosome analysis.\u00a0\u00a0FISH is the ideal choice for detection of this deletion due to its increased sensitivity.<\/p>\n

Methods<\/strong><\/p>\n

The sample is usually cultured until sufficient number of mitotic cells is observed (48-72 hours for peripheral blood and 7-10 days for amniotic fluid) and after harvesting the cells are hybridized with the probes.\u00a0\u00a0After hybridization and post-hybridization washes, the slides are analyzed under a fluorescence microscope equipped with\u00a0epifluorescence\u00a0and appropriate filter sets.\u00a0\u00a0Analysis and scoring is carried out by two certified technologists in a blinded fashion with each scoring a minimum of 5 metaphases and 50 interphase nuclei.\u00a0\u00a0Results are reviewed by both the laboratory manager and the director.<\/p>\n

15q11 deletion (Prader-Willi syndrome)\u00a0SNRPN \/ 154P1\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0Cytocell\u00a0(cat # LPU005)<\/p>\n

Interpretation of Results<\/strong><\/p>\n

The morphologic interpretation and correlation of results on all cases is performed by a board-certified doctoral level scientist (laboratory director).\u00a0The final report has a narrative description of results.\u00a0\u00a0If abnormalities are present, they are explained in a paragraph which helps to clarify and correlate chromosomal findings with phenotype.\u00a0References are included in the report to help the referring physician with interpretation, which include books or journals that contain appropriate information. Recommendations are made as to any additional testing, if necessary.<\/p>\n

[\/vc_column_text][\/vc_column][vc_column width=”1\/2″][vc_column_text]<\/p>\n

Angelman Syndrome<\/h3>\n

Test Overview<\/p>\n

CPT Code(s)<\/strong><\/p>\n

Peripheral Blood: 88230-52, 88271, 88273, 88291<\/p>\n

Amniotic Fluid: 88235-52, 88271, 88273, 88291<\/p>\n

Methodology<\/strong><\/p>\n

Culture\/Hybridization\/Microscopy\/Interpretation<\/p>\n

Specimen Requirements<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n
<\/td>\nPeripheral Blood<\/strong><\/td>\nAmniotic Fluid<\/strong><\/td>\n<\/tr>\n
Container<\/td>\nSodium Heparin (green top tube)<\/td>\nTwo 15 ml sterile leak-proof conical tubes<\/td>\n<\/tr>\n
Optimal Quantity<\/td>\n2-3 ml<\/td>\n20-25 ml<\/td>\n<\/tr>\n
Minimum Quantity<\/td>\n1-2 ml<\/td>\n10 ml<\/td>\n<\/tr>\n
Storage<\/td>\nRoom Temperature<\/td>\nRoom Temperature<\/td>\n<\/tr>\n
Stability at Room Temperature<\/td>\nRoom Temperature for 8 hours, then refrigerate<\/td>\nRoom Temperature for 8 hours, then refrigerate<\/td>\n<\/tr>\n
Transportation<\/td>\nAvoid freezing or heating over 35o<\/sup>C<\/td>\nAvoid freezing or heating over 35o<\/sup>C<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

Turnaround Time<\/strong><\/p>\n

Peripheral Blood: Final report in 5 days for 90% of cases<\/p>\n

Amniotic Fluid: Final report in 7-10 days for 90% of cases<\/p>\n

Test Details<\/p>\n

Clinical Significance<\/strong><\/p>\n

Fluorescence in situ hybridization (FISH) is a sensitive method to detect smaller genomic changes associated with various\u00a0micordeletion\u00a0syndromes.\u00a0\u00a0There are several advantages with FISH technology over routine chromosome analysis and such advantages include the ability of FISH technology to detect genomic abnormalities in non-viable and non-dividing tissues, rapid turnaround time, and increased resolution.\u00a0\u00a0However, FISH technology is complementary to routine chromosome analysis and cannot substitute routine chromosome analysis and FISH diagnostic testing is\u00a0nor\u00a0recommended as an isolated test without chromosome analysis for the diagnosis of these\u00a0microdeletion\u00a0syndromes.<\/p>\n

Clinical Background<\/strong><\/p>\n

Angelman\u00a0syndrome, caused by\u00a0microdeletions\/loss of maternally derived gene expression, is a\u00a0panethnic\u00a0developmental disorder affecting approximately 1 in 10,000 to 1 in 15,000 live births. Patients with this syndrome show characteristic features including unusual facial appearance, short stature, severe mental retardation, spasticity and seizures.\u00a0\u00a0Chromosome deletion 15q11q13 during female meiosis results in Angelman syndrome and such deletions account for a large number of patients while a small number of patients have mutations within this gene that cannot be detected by either routine chromosome analysis or FISH.\u00a0\u00a0\u00a0Since the most common deletion encompasses ~3 Mb, often these deletions are not detected on routine chromosome analysis.\u00a0\u00a0FISH is the ideal choice for detection of this deletion due to its increased sensitivity.<\/p>\n

Methods<\/strong><\/p>\n

The sample is usually cultured until sufficient number of mitotic cells is observed (48-72 hours for peripheral blood and 7-10 days for amniotic fluid) and after harvesting the cells are hybridized with the probes.\u00a0\u00a0After hybridization and post-hybridization washes, the slides analyzed under a fluorescence microscope equipped with\u00a0epifluorescence\u00a0and appropriate filter sets.\u00a0\u00a0Analysis and scoring is carried out by two certified technologists in a blinded fashion with each scoring a minimum of 5 metaphases and 50 interphase nuclei.\u00a0\u00a0Results are reviewed by both the laboratory manager and the director.<\/p>\n

15q11 deletion (Angelman\u00a0syndrome)\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0UBE3A \/ 154P1\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0Cytocell\u00a0(cat # LPU006)<\/p>\n

Interpretation of Results<\/strong><\/p>\n

The morphologic interpretation and correlation of results on all cases is performed by a board-certified doctoral level scientist (laboratory director).\u00a0The final report has a narrative description of results.\u00a0\u00a0If abnormalities are present, they are explained in a paragraph which helps to clarify and correlate chromosomal findings with phenotype.\u00a0References are included in the report to help the referring physician with interpretation, which include books or journals that contain appropriate information. Recommendations are made as to any additional testing, if necessary.<\/p>\n

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