Cholestasis Sequencing Panel

  • Summary and Pricing
  • Clinical Features and Genetics
  • Citations
  • Methods
  • Ordering/Specimens
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NGS Sequencing

Test Code Test Copy GenesCPT Code Copy CPT Codes
1963 ABCB11 81479 Add to Order
ABCB4 81479
AKR1D1 81479
ATP8B1 81479
BAAT 81479
CLDN1 81479
HSD3B7 81479
JAG1 81407
NOTCH2 81479
NR1H4 81479
SERPINA1 81479
SLC25A13 81479
TJP2 81479
VIPAS39 81479
VPS33B 81479
Full Panel Price* $1590.00
Test Code Test Copy Genes Total Price CPT Codes Copy CPT Codes
1963 Genes x (15) $1590.00 81407, 81479(x14) Add to Order
Pricing Comment

If you would like to order a subset of these genes contact us to discuss pricing.

Targeted Testing

For ordering targeted known variants, please proceed to our Targeted Variants landing page.

Turnaround Time

The great majority of tests are completed within 28 days.

Clinical Sensitivity

Detection rate of pathogenic variants in each gene of this panel in a large cohort of patients with cholestasis of undefined etiology is unknown. In a study of 51 subjects with cholestasis of undefined etiology, Matte et al. found causative variants in JAG1, ATP8B1, ABCB11, or ABCB4 genes in 14 patients (27%) (Matte et al. 2010). ATP8B1 and ABCB11 causative variants were found in 12 (21%) and 36 (63%) respectively of 57 families affected by PFIC with normal GGT activity (Davit-Spraul et al. 2010). In a cohort study of 68 unrelated PFIC3 patients, ABCB4 causative variants were found at both alleles in 13 patients (19%) and only one allele in 5 patients (7%), respectively (Degiorgio et al. 2007). Out of 29 families that had chronic cholestatic liver disease with low GGT and did not have causative variants in ABCB11 or ATP8B1, TJP2 mutations were found in 8 families (27.6%) (Sambrotta et al. 2014).

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Deletion/Duplication Testing via aCGH

Test Code Test Copy GenesIndividual Gene PriceCPT Code Copy CPT Codes
600 ABCB11$690.00 81479 Add to Order
ABCB4$690.00 81479
AKR1D1$690.00 81479
ATP8B1$690.00 81479
BAAT$690.00 81479
CLDN1$690.00 81479
HSD3B7$690.00 81479
JAG1$690.00 81406
NOTCH2$690.00 81479
NR1H4$690.00 81479
SERPINA1$690.00 81479
SLC25A13$690.00 81479
TJP2$690.00 81479
VIPAS39$690.00 81479
VPS33B$690.00 81479
Full Panel Price* $1290.00
Test Code Test Copy Genes Total Price CPT Codes Copy CPT Codes
600 Genes x (15) $1290.00 81406, 81479(x14) Add to Order
Pricing Comment

# of Genes Ordered

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Over 100

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Turnaround Time

The great majority of tests are completed within 28 days.

Clinical Sensitivity

The prevalence of large deletions (vast majority) and duplications affecting the JAG1 gene are expected to be about 5% in Alagille syndrome (Warthen et al. 2006). To date, only two large deletions have been found to affect the VPS33B gene, accounting for 4% of total documented pathogenic variants in this gene (Human Gene Mutation Database). No gross deletions or duplications have been reported in VIPAS39. Large deletions and duplications (more often) have been found in the SLC25A13 locus, accounting for 7% of all documented pathogenic variants in this gene (Human Gene Mutation Database). In a study of 68 East Asian patients affected by citrin deficiency (CD), one patient (1.5%) was found to have a large SLC25A13 duplication (Song et al. 2013).

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Clinical Features

Cholestasis is characterized by jaundice and pruritus. It can present as the hallmark feature in progressive familial intrahepatic cholestasis (PFIC) or as a feature in other inherited disorders such as Alagille syndrome.PFIC is a group of autosomal recessive liver disorders due to defects in bile secretion and is characterized by intrahepatic cholestasis with disease onset usually in infancy and childhood (Srivastava 2014; Davit-Spraul et al. 2009). Four types of PFIC have been classified in terms of causative genes involved in the hepatocellular transport system. FIC1 (familial intrahepatic cholestasis protein 1 deficiency (PFIC1) is caused by mutations in ATP8B1, and BSEP (bile salt export pump). PFIC2 is caused by mutations in ABCB11. PFIC3 is caused by reduced biliary phospholipid secretion due to mutations in ABCB4. The most recently identified PFIC4 is caused by abnormal tight junctions between adjacent hepatocytes and biliary canaliculi in liver tissue due to TJP2 defects (Sambrotta et al. 2014). PFIC patients usually develop fibrosis and end-stage liver disease before adulthood. Serum gamma-glutamyl transferase (GGT) activity is normal in PFIC1 and PFIC2 patients, elevated in PFIC3 patients and low in PFIC4 patients.Defects in PFIC-associated genes ATP8B1 and ABCB11 can also cause a milder type of liver disease called benign recurrent intrahepatic cholestasis (BRIC) (Klomp et al. 2004; Beauséjour et al. 2011). BRIC is an autosomal recessive disorder characterized by intermittent episodes of cholestasis without progression to liver failure.Other inherited disorders featuring cholestasis includes Alagille syndrome (JAG1 and NOTCH2), arthrogryposis, renal dysfunction, and cholestasis syndrome (ARC syndrome; VPS33B and VIPAS39), alpha-1-antitrypsin deficiency (SERPINA1), citrullinemia (SLC25A13), congenital defects of bile acid synthesis (HSD3B7 and AKR1D1), familial hypercholanemia (TJP2 and BAAT) and neonatal ichthyosis-sclerosing cholangitis syndrome (CLDN1). More information about each gene can be found in the Genetics section below.


Progressive familial intrahepatic cholestasis (PFIC) is inherited in an autosomal recessive manner. Defects in four genes to date encoding proteins associated with hepatocellular transport system have been found to cause PFIC: ATP8B1, ABCB11, ABCB4 and TJP2 (Srivastava et al. 2014; Davit-Spraul et al. 2009; Sambrotta et al. 2014) (see also Clinical Features above). The ATP8B1 gene (27 coding exons) encodes the FIC1 protein, which is a member of the P-type cation transport ATPase family. Pathogenic defects in ATP8B1 include missense, nonsense, splicing site mutations, small indels and exon-level large deletions (Human Gene Mutation Database). In addition to PFIC1, recessive ATP8B1 mutations can also cause benign recurrent intrahepatic cholestasis-1 (BRIC1). The ABCB11 (27 coding exons) gene encodes the bile salt export pump protein (BSEP), which is a member of the superfamily of ATP-binding cassette (ABC) transporters. Pathogenic defects in ABCB11 include missense, nonsense, splicing site mutations and small indels and exon-level large deletions (Human Gene Mutation Database). In addition to PFIC2, recessive ABCB11 mutations can also cause benign recurrent intrahepatic cholestasis-2 (BRIC2). The ABCB4 gene (27 coding exons) encodes the multi-drug resistant 3 protein (MDR3), which is also a member of the superfamily of ATP-binding cassette (ABC) transporters. Pathogenic defects in ABCB4 include missense, nonsense, splicing site mutations, small indels and exon-level large deletions (Human Gene Mutation Database). The newly found fourth PFIC gene TJP2 (23 coding exons) encodes the tight junction protein 2, which is involved in the organization of epithelial and endothelial intercellular junctions. Pathogenic defects in TJP2 include missense, nonsense, splicing site mutations, small indels and exon-level large deletions (Human Gene Mutation Database). TJP2 defects can also cause familial hypercholanemia, which is characterized by elevated serum bile acid concentrations, itching, and fat malabsorption (Carlton et al. 2003).

Genes associated with other inherited disorders featuring cholestasis are described as follows.The JAG1 gene (26 coding exons) encodes the ligand for the receptor Notch 1 in the Notch signaling pathway. Dominant JAG1 mutations cause Alagille syndrome (Oda et al. 1997; Warthen et al. 2006) (see Test # 427).The NOTCH2 gene (34 coding exons) encodes the receptor Notch 2 in the Notch signaling pathway. Dominant NOTCH2 mutations can cause both Alagille syndrome (McDaniell et al. 2006) and the skeletal disorder Hajdu-Cheney syndrome (Simpson et al. 2011).The VPS33B gene (23 coding exons) encodes a homolog of the yeast vacuolar sorting protein Vps33, which is a cytosolic protein involved in vesicle membrane fusion. Recessive VPS33B mutations are the major causes for arthrogryposis, renal dysfunction, and cholestasis syndrome (ARC syndrome) (Gissen et al. 2004) (see Test # 248).The VIPAS39 gene (21 coding exons) encodes the VPS33B interacting protein, apical-basolateral polarity regulator (VIPAR). Recessive VIPAS39 mutations are a less common cause for arthrogryposis, renal dysfunction, and cholestasis syndrome (ARC syndrome) (Cullinane et al. 2010) (see Test # 591).The SERPINA1 gene (4 coding exons) encodes alpha-1-antitrypsin (AAT), also known as protease inhibitor (PI), a major plasma serine protease inhibitor. Recessive SERPINA1 mutations cause alpha-1-antitrypsin deficiency (Medicina et al. 2009).The SLC25A13 gene (18 coding exons) encodes citrin, a calcium-dependent mitochondrial solute transporter with a role in urea cycle function. Recessive SLC25A13 mutations can cause both adult-onset type II citrullinemia (Kobayashi et al. 1999) and neonatal-onset type II citrullinemia (Tazawa et al. 2004).The HSD3B7 gene (7 coding exons) encodes 3 beta-hydroxysteroid dehydrogenase type 7, which is an enzyme in the production of bile acids. Recessive HSD3B7 mutations cause congenital bile acid synthesis defect type 1 (CBAS1) (Cheng et al. 2003).The AKR1D1 gene (9 coding exons) encodes the steroid 5-beta-reductase, which is responsible for catalysis of the 5-beta-reduction of bile acid intermediates and steroid hormones carrying a delta (4)-3-one structure. Recessive AKR1D1 mutations cause congenital bile acid synthesis defect type 2 (CBAS2) (Lemonde et al. 2003).The NR1H4 gene (9 coding exons) encodes a ligand-activated transcription factor, which functions as a receptor for bile acids. A heterozygous nonsense NR1H4 mutation has been reported in idiopathic infantile cholestasis (Chen et al. 2012).The BAAT gene (3 coding exons) encodes bile acid-CoA:amino acid N-acyltransferase, a liver enzyme that catalyzes the transfer of C24 bile acids from the acyl-CoA thioester to either glycine or taurine. Recessive BAAT mutations cause bile acid amidation defect (Setchell et al. 2013).The CLDN1 gene (4 coding exons) encodes a member of the claudin family, which is an integral membrane protein and a component of tight junction strands. Recessive CLDN1 mutations result in neonatal ichthyosis-sclerosing cholangitis syndrome (NISCH) (Feldmeyer et al. 2006).

Testing Strategy

This NextGen Sequencing Panel analyzes 16 genes that have been associated with cholestasis. For this NGS panel, the full coding regions, plus ~20 bp of non-coding DNA flanking each exon, are sequenced for each of the genes listed below. Sequencing is accomplished by capturing specific regions with an optimized solution-based hybridization method, followed by massively parallel sequencing of the captured DNA fragments. Additional Sanger sequencing is performed for any regions not captured or with insufficient number of sequence reads. All pathogenic, undocumented and questionable variant calls are confirmed by Sanger sequencing.

This test does not currently include sequencing or gene-centric aCGH for exons 1 to 4 of the NOTCH2 gene because of high sequence similarity to one or more additional chromosomal regions. So far, no pathogenic variants have been reported in these exons.

Indications for Test

Candidates for this test are patients with cholestasis. This test especially aids in a differential diagnosis of similar phenotypes by analyzing multiple genes simultaneously.


Official Gene Symbol OMIM ID
ABCB11 603201
ABCB4 171060
AKR1D1 604741
ATP8B1 602397
BAAT 602938
CLDN1 603718
HSD3B7 607764
JAG1 601920
NOTCH2 600275
NR1H4 603826
SERPINA1 107400
SLC25A13 603859
TJP2 607709
VIPAS39 613401
VPS33B 608552
Inheritance Abbreviation
Autosomal Dominant AD
Autosomal Recessive AR
X-Linked XL
Mitochondrial MT


Name Inheritance OMIM ID
Alagille Syndrome 1 118450
Alagille Syndrome 2 610205
Alpha-1-Antitrypsin Deficiency 613490
Arthrogryposis Renal Dysfunction Cholestasis Syndrome 208085
Arthrogryposis, Renal Dysfunction, And Cholestasis 2 613404
Benign Recurrent Intrahepatic Cholestasis 1 243300
Benign Recurrent Intrahepatic Cholestasis 2 605479
Bile Acid Synthesis Defect, Congenital, 1 607765
Bile Acid Synthesis Defect, Congenital, 2 235555
Cholestasis, Progressive Familial Intrahepatic 2 601847
Cholestasis, Progressive Familial Intrahepatic 3 602347
Cholestasis, Progressive Familial Intrahepatic 4 615878
Citrin Deficiency 605814
Citrullinemia Type II 603471
Hajdu-Cheney Syndrome 102500
Hypercholanemia, Familial 607748
Ichthyosis, Leukocyte Vacuoles, Alopecia, And Sclerosing Cholangitis 607626
Progressive Intrahepatic Cholestasis 211600

Related Tests

NOTCH2-Related Disorders via the NOTCH2 Gene
SLC25A13-Related Disorders via the SLC25A13 Gene
Alagille Syndrome Sequencing Panel
Alagille Syndrome-1 via the JAG1 Gene
Alpha-1 Antitrypsin Deficiency via the SERPINA1 Gene
Arthrogryposis-Renal Dysfunction-Cholestasis (ARC) Syndrome via the VIPAS39 Gene
Arthrogryposis-Renal Dysfunction-Cholestasis (ARC) Syndrome via the VPS33B Gene
Autism Spectrum Disorders and Intellectual Disability (ASD-ID) Comprehensive Sequencing Panel with CNV Detection
Congenital Bile Acid Synthesis Defect Type 1 via the HSD3B7 Gene
Congenital Bile Acid Synthesis Defect Type 2 via the AKR1D1 Gene
Congenital Ichthyosis and Related Disorders Sequencing Panel with CNV Detection
Deafness, Autosomal Dominant 51 (DFNA51) via the TJP2 Gene
Hyperammonemia Sequencing Panel
Intrahepatic Cholestasis via the ABCB11 Gene
Intrahepatic Cholestasis via the ABCB4 Gene
Intrahepatic Cholestasis via the ATP8B1 Gene
Nonsyndromic Hearing Loss and Deafness Sequencing Panel
Progressive Familial Intrahepatic Cholestasis (PFIC) and Alagille syndrome Sequencing Panel
Urea Cycle Disorders Sequencing Panel


Genetic Counselors
  • Beauséjour Y. et al. 2011. Canadian Journal of Gastroenterology = Journal Canadien De Gastroenterologie. 25: 311-4. PubMed ID: 21766090
  • Carlton VEH, Harris BZ, Puffenberger EG, Batta AK, Knisely AS, Robinson DL, Strauss KA, Shneider BL, Lim WA, Salen G, Morton DH, Bull LN. 2003. Complex inheritance of familial hypercholanemia with associated mutations in TJP2 and BAAT. Nat. Genet. 34: 91–96. PubMed ID: 12704386
  • Chen X-Q, Wang L-L, Shan Q-W, Tang Q, Deng Y-N, Lian S-J, Yun X. 2012. A novel heterozygous NR1H4 termination codon mutation in idiopathic infantile cholestasis. World J Pediatr 8: 67–71. PubMed ID: 21633855
  • Cheng JB, Jacquemin E, Gerhardt M, Nazer H, Cresteil D, Heubi JE, Setchell KDR, Russell DW. 2003. Molecular genetics of 3beta-hydroxy-Delta5-C27-steroid oxidoreductase deficiency in 16 patients with loss of bile acid synthesis and liver disease. J. Clin. Endocrinol. Metab. 88: 1833–1841. PubMed ID: 12679481
  • Cullinane AR, Straatman-Iwanowska A, Zaucker A, Wakabayashi Y, Bruce CK, Luo G, Rahman F, Gürakan F, Utine E, Ozkan TB, Denecke J, Vukovic J, Di Rocco M, Mandel H, Cangul H, Matthews RP, Thomas SG, Rappoport JZ, Arias IM, Wolburg H, Knisely AS, Kelly DA, Müller F, Maher ER, Gissen P. 2010. Mutations in VIPAR cause an arthrogryposis, renal dysfunction and cholestasis syndrome phenotype with defects in epithelial polarization. Nat. Genet. 42: 303–312. PubMed ID: 20190753
  • Davit-Spraul A. et al. 2009. Orphanet Journal of Rare Diseases. 4: 1. PubMed ID: 19133130
  • Davit-Spraul A. et al. 2010. Hepatology (baltimore, Md.). 51: 1645-55. PubMed ID: 20232290
  • Degiorgio D. et al. 2007. European Journal of Human Genetics : Ejhg. 15: 1230-8. PubMed ID: 17726488
  • Feldmeyer L, Huber M, Fellmann F, Beckmann JS, Frenk E, Hohl D. 2006. Confirmation of the origin of NISCH syndrome. Hum. Mutat. 27: 408–410. PubMed ID: 16619213
  • Gissen P, Johnson CA, Morgan NV, Stapelbroek JM, Forshew T, Cooper WN, McKiernan PJ, Klomp LWJ, Morris AAM, Wraith JE, McClean P, Lynch SA, Thompson RJ, Lo B, Quarrell OW, Di Rocco M, Trembath RC, Mandel H, Wali S, Karet FE, Knisely AS, Houwen RH, Kelly DA, Maher ER. 2004. Mutations in VPS33B, encoding a regulator of SNARE-dependent membrane fusion, cause arthrogryposis-renal dysfunction-cholestasis (ARC) syndrome. Nat. Genet. 36: 400–404.  PubMed ID: 15052268
  • Human Gene Mutation Database (Bio-base).
  • Klomp L.W. et al. 2004. Hepatology (baltimore, Md.). 40: 27-38. PubMed ID: 15239083
  • Kobayashi K, Sinasac DS, Iijima M, Boright AP, Begum L, Lee JR, Yasuda T, Ikeda S, Hirano R, Terazono H, Crackower MA, Kondo I, Tsui LC, Scherer SW, Saheki T. 1999. The gene mutated in adult-onset type II citrullinaemia encodes a putative mitochondrial carrier protein. Nat. Genet. 22: 159–163. PubMed ID: 10369257
  • Lemonde H.A. et al. 2003. Gut. 52: 1494-9. PubMed ID: 12970144
  • Matte U. et al. 2010. Journal of Pediatric Gastroenterology and Nutrition. 51: 488-93. PubMed ID: 20683201
  • McDaniell R, Warthen DM, Sanchez-Lara PA, Pai A, Krantz ID, Piccoli DA, Spinner NB. 2006. NOTCH2 mutations cause Alagille syndrome, a heterogeneous disorder of the notch signaling pathway. Am. J. Hum. Genet. 79: 169–173. PubMed ID: 16773578
  • Medicina D, Montani N, Fra AM, Tiberio L, Corda L, Miranda E, Pezzini A, Bonetti F, Ingrassia R, Scabini R, Facchetti F, Schiaffonati L. 2009. Molecular characterization of the new defective P(brescia) alpha1-antitrypsin allele. Hum. Mutat. 30: E771–781. PubMed ID: 19437508
  • Oda T, Elkahloun AG, Pike BL, Okajima K, Krantz ID, Genin A, Piccoli DA, Meltzer PS, Spinner NB, Collins FS, Chandrasekharappa SC. 1997. Mutations in the human Jagged1 gene are responsible for Alagille syndrome. Nat. Genet. 16: 235–242. PubMed ID: 9207787
  • Sambrotta M. et al. 2014. Nature Genetics. 46: 326-8. PubMed ID: 24614073
  • Setchell KDR, Heubi JE, Shah S, Lavine JE, Suskind D, Al–Edreesi M, Potter C, Russell DW, O’Connell NC, Wolfe B, Jha P, Zhang W, Bove KE, Knisely AS, Hofmann AF, Rosenthal P, Bull LN. 2013. Genetic Defects in Bile Acid Conjugation Cause Fat-Soluble Vitamin Deficiency. Gastroenterology 144: 945–955.e6. PubMed ID: 23415802
  • Simpson MA, Irving MD, Asilmaz E, Gray MJ, Dafou D, Elmslie FV, Mansour S, Holder SE, Brain CE, Burton BK, Kim KH, Pauli RM, Aftimos S, Stewart H, Kim CA, Holder-Espinasse M, Robertson SP, Drake WM, Trembath RC. 2011. Mutations in NOTCH2 cause Hajdu-Cheney syndrome, a disorder of severe and progressive bone loss. Nat. Genet. 43: 303–305. PubMed ID: 21378985
  • Song Y-Z, Zhang Z-H, Lin W-X, Zhao X-J, Deng M, Ma Y-L, Guo L, Chen F-P, Long X-L, He X-L, Sunada Y, Soneda S, Nakatomi A, Dateki S, Ngu LH, Kobayashi K, Saheki T. 2013. SLC25A13 Gene Analysis in Citrin Deficiency: Sixteen Novel Mutations in East Asian Patients, and the Mutation Distribution in a Large Pediatric Cohort in China. PLoS ONE 8: e74544. PubMed ID: 24069319
  • Srivastava A. 2014. Journal of Clinical and Experimental Hepatology. 4: 25-36. PubMed ID: 25755532
  • Tazawa Y, Kobayashi K, Abukawa D, Nagata I, Maisawa S, Sumazaki R, Iizuka T, Hosoda Y, Okamoto M, Murakami J, Kaji S, Tabata A, Lu YB, Sakamoto O, Matsui A, Kanzaki S, Takada G, Saheki T, Iinuma K, Ohura T. 2004. Clinical heterogeneity of neonatal intrahepatic cholestasis caused by citrin deficiency: case reports from 16 patients. Mol. Genet. Metab. 83: 213–219. PubMed ID: 15542392
  • Warthen DM, Moore EC, Kamath BM, Morrissette JJD, Sanchez-Lara PA, Sanchez P, Piccoli DA, Krantz ID, Spinner NB. 2006. Jagged1 (JAG1) mutations in Alagille syndrome: increasing the mutation detection rate. Hum. Mutat. 27: 436–443. PubMed ID: 16575836
Order Kits

NextGen Sequencing using PG-Select Capture Probes

Test Procedure

We use a combination of Next Generation Sequencing (NGS) and Sanger sequencing technologies to cover the full coding regions of the listed genes plus ~20 bases of non-coding DNA flanking each exon.  As required, genomic DNA is extracted from the patient specimen.  For NGS, patient DNA corresponding to these regions is captured using an optimized set of DNA hybridization probes.  Captured DNA is sequenced using Illumina’s Reversible Dye Terminator (RDT) platform (Illumina, San Diego, CA, USA).  Regions with insufficient coverage by NGS are covered by Sanger sequencing.  All pathogenic, likely pathogenic, or variants of uncertain significance are confirmed by Sanger sequencing.

For Sanger sequencing, Polymerase Chain Reaction (PCR) is used to amplify targeted regions.  After purification of the PCR products, cycle sequencing is carried out using the ABI Big Dye Terminator v.3.0 kit.  PCR products are resolved by electrophoresis on an ABI 3730xl capillary sequencer.  In nearly all cases, cycle sequencing is performed separately in both the forward and reverse directions.

Patient DNA sequence is aligned to the genomic reference sequence for the indicated gene region(s). All differences from the reference sequences (sequence variants) are assigned to one of five interpretation categories, listed below, per ACMG Guidelines (Richards et al. 2015).

(1) Pathogenic Variants
(2) Likely Pathogenic Variants
(3) Variants of Uncertain Significance
(4) Likely Benign Variants
(5) Benign, Common Variants

Human Genome Variation Society (HGVS) recommendations are used to describe sequence variants (  Rare variants and undocumented variants are nearly always classified as likely benign if there is no indication that they alter protein sequence or disrupt splicing.

Analytical Validity

As of March 2016, 6.36 Mb of sequence (83 genes, 1557 exons) generated in our lab was compared between Sanger and NextGen methodologies. We detected no differences between the two methods. The comparison involved 6400 total sequence variants (differences from the reference sequences). Of these, 6144 were nucleotide substitutions and 256 were insertions or deletions. About 65% of the variants were heterozygous and 35% homozygous. The insertions and deletions ranged in length from 1 to over 100 nucleotides.

In silico validation of insertions and deletions in 20 replicates of 5 genes was also performed. The validation included insertions and deletions of lengths between 1 and 100 nucleotides. Insertions tested in silico: 2200 between 1 and 5 nucleotides, 625 between 6 and 10 nucleotides, 29 between 11 and 20 nucleotides, 25 between 21 and 49 nucleotides, and 23 at or greater than 50 nucleotides, with the largest at 98 nucleotides. All insertions were detected. Deletions tested in silico: 1813 between 1 and 5 nucleotides, 97 between 6 and 10 nucleotides, 32 between 11 and 20 nucleotides, 20 between 21 and 49 nucleotides, and 39 at or greater than 50 nucleotides, with the largest at 96 nucleotides. All deletions less than 50 nucleotides in length were detected, 13 greater than 50 nucleotides in length were missed. Our standard NextGen sequence variant calling algorithms are generally not capable of detecting insertions (duplications) or heterozygous deletions greater than 100 nucleotides. Large homozygous deletions appear to be detectable.   

Analytical Limitations

Interpretation of the test results is limited by the information that is currently available.  Better interpretation should be possible in the future as more data and knowledge about human genetics and this specific disorder are accumulated.

When Sanger sequencing does not reveal any difference from the reference sequence, or when a sequence variant is homozygous, we cannot be certain that we were able to detect both patient alleles.  Occasionally, a patient may carry an allele which does not amplify, due to a large deletion or insertion.   In these cases, the report will contain no information about the second allele.  Our Sanger and NGS Sequencing tests are generally not capable of detecting Copy Number Variants (CNVs).

We sequence all coding exons for each given transcript, plus ~20 bp of flanking non-coding DNA for each exon.  Test reports contain no information about other portions of the gene, such as regulatory domains, deep intronic regions or any currently uncharacterized alternative exons.

In most cases, we are unable to determine the phase of sequence variants.  In particular, when we find two likely causative mutations for recessive disorders, we cannot be certain that the mutations are on different alleles.

Our ability to detect minor sequence variants due to somatic mosaicism is limited.  Sequence variants that are present in less than 50% of the patient’s nucleated cells may not be detected.

Runs of mononucleotide repeats (eg (A)n or (T)n) with n >8 in the reference sequence are generally not analyzed because of strand slippage during PCR.

Unless otherwise indicated, DNA sequence data is obtained from a specific cell-type (usually leukocytes from whole blood).   Test reports contain no information about the DNA sequence in other cell-types.

We cannot be certain that the reference sequences are correct.

Rare, low probability interpretations of sequencing results, such as for example the occurrence of de novo mutations in recessive disorders, are generally not included in the reports.

We have confidence in our ability to track a specimen once it has been received by PreventionGenetics.  However, we take no responsibility for any specimen labeling errors that occur before the sample arrives at PreventionGenetics.

Deletion/Duplication Testing Via Array Comparative Genomic Hybridization

Test Procedure

Equal amounts of genomic DNA from the patient and a gender matched reference sample are amplified and labeled with Cy3 and Cy5 dyes, respectively. To prevent any sample cross contamination, a unique sample tracking control is added into each patient sample. Each labeled patient product is then purified, quantified, and combined with the same amount of reference product. The combined sample is loaded onto the designed array and hybridized for at least 22-42 hours at 65°C. Arrays are then washed and scanned immediately with 2.5 µM resolution. Only data for the gene(s) of interest for each patient are extracted and analyzed.

Analytical Validity

PreventionGenetics' high density gene-centric custom designed aCGH enables the detection of relatively small deletions and duplications within a single exon of a given gene or deletions and duplications encompassing the entire gene. PreventionGenetics has established and verified this test's accuracy and precision.

Analytical Limitations

Our dense probe coverage may allow detection of deletions/duplications down to 100 bp; however due to limitations and probe spacing this cannot be guaranteed across all exons of all genes. Therefore, some copy number changes smaller than 100-300 bp within a targeted large exon may not be detected by our array.

This array may not detect deletions and duplications present at low levels of mosaicism or those present in genes that have pseudogene copies or repeats elsewhere in the genome.

aCGH will not detect balanced translocations, inversions, or point mutations that may be responsible for the clinical phenotype.

Breakpoints, if occurring outside the targeted gene, may be hard to define.

The sensitivity of this assay may be reduced when DNA is extracted by an outside laboratory.

Order Kits

Ordering Options

myPrevent - Online Ordering
  • The test can be added to your online orders in the Summary and Pricing section.
  • Once the test has been added log in to myPrevent to fill out an online requisition form.
  • A completed requisition form must accompany all specimens.
  • Billing information along with specimen and shipping instructions are within the requisition form.
  • All testing must be ordered by a qualified healthcare provider.


(Delivery accepted Monday - Saturday)

  • Collect 3 ml -5 ml (5 ml preferred) of whole blood in EDTA (purple top tube) or ACD (yellow top tube). For Test #500-DNA Banking only, collect 10 ml -20 ml of whole blood.
  • For small babies, we require a minimum of 1 ml of blood.
  • Only one blood tube is required for multiple tests.
  • Ship blood tubes at room temperature in an insulated container. Do not freeze blood.
  • During hot weather, include a frozen ice pack in the shipping container. Place a paper towel or other thin material between the ice pack and the blood tube.
  • In cold weather, include an unfrozen ice pack in the shipping container as insulation.
  • At room temperature, blood specimen is stable for up to 48 hours.
  • If refrigerated, blood specimen is stable for up to one week.
  • Label the tube with the patient name, date of birth and/or ID number.


(Delivery accepted Monday - Saturday)

  • Send in screw cap tube at least 5 µg -10 µg of purified DNA at a concentration of at least 20 µg/ml for NGS and Sanger tests and at least 5 µg of purified DNA at a concentration of at least 100 µg/ml for gene-centric aCGH, MLPA, and CMA tests, minimum 2 µg for limited specimens.
  • For requests requiring more than one test, send an additional 5 µg DNA per test ordered when possible.
  • DNA may be shipped at room temperature.
  • Label the tube with the composition of the solute, DNA concentration as well as the patient’s name, date of birth, and/or ID number.
  • We only accept genomic DNA for testing. We do NOT accept products of whole genome amplification reactions or other amplification reactions.


(Delivery preferred Monday - Thursday)

  • PreventionGenetics should be notified in advance of arrival of a cell culture.
  • Culture and send at least two T25 flasks of confluent cells.
  • Some panels may require additional flasks (dependent on size of genes, amount of Sanger sequencing required, etc.). Multiple test requests may also require additional flasks. Please contact us for details.
  • Send specimens in insulated, shatterproof container overnight.
  • Cell cultures may be shipped at room temperature or refrigerated.
  • Label the flasks with the patient name, date of birth, and/or ID number.
  • We strongly recommend maintaining a local back-up culture. We do not culture cells.
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