Use of a Comprehensive 66-Gene Cholestasis Sequencing Panel in 2171 Cholestatic Infants, Children, and Young Adults.

OBJECTIVES: Cholestasis is caused by a wide variety of etiologies, often genetic in origin. Broad overlap in clinical presentations, particularly in newborns, renders prioritizing diagnostic investigations challenging. In this setting, a timely, comprehensive assessment using a multigene panel by a clinical diagnostic laboratory would likely prove useful. We summarize initial findings from a testing program designed to discover genetic causes of cholestasis. METHODS: A neonatal/adult sequencing panel containing 66 genes (originally 57; nine added March 2017) relevant to cholestasis was used. A broad range of eligible patients were enrolled with current/history of cholestasis without an identified cause, or unexplained chronic liver disease. DNA sequencing utilized a custom-designed capture library, and variants were classified and reported as benign, likely benign, variant of unknown significance (VOUS), likely pathogenic (LP), or pathogenic (P), according to the clinical interpretation workflow at EGL Genetics (Tucker, GA). RESULTS: A total of 2433 samples were submitted between February 2016 and December 2017; 2171 results were reported. Median turnaround time was 21 days. Results from the 2171 subjects (57% <1 year old) included 583 P variants, 79 LP variants, and 3117 VOUS; 166 P/LP variants and 415 VOUS were novel. The panel's overall diagnostic yield was 12% (n = 265/2171) representing 32 genes. The top five genetic diagnoses for the group, in order: JAG1 + NOTCH2 (Alagille syndrome), ABCB11, SERPINA1, ABCB4, and POLG. CONCLUSIONS: These findings support the utility of comprehensive rapid multigene testing in diagnosing cholestasis and highlight the evolving understanding of genetic variants contributing to the pathogenesis of cholestasis.

Calibration and validation of the 2WIN photoscreener compared to the PlusoptiX S12 and the SPOT.

PURPOSE: Pediatricians are interested in the amblyopia detection ability of photoscreeners, whereas ophthalmologists ponder their value as autorefractors. The 2WIN (Adaptica, Padova, Italy) is a new device capable of estimating refractive error and ocular alignment by infrared photoscreening. METHODS: Sequential pediatric eye patients with a high (56% to 60%) prescreening prevalence of amblyopia risk factors were screened with the PlusoptiX S12 (PlusoptiX, Inc., Atlanta, GA), SPOT (PediaVision, Lake Mary, FL), and 2WIN photoscreeners before confirmatory examination adhering to American Association for Pediatric Ophthalmology and Strabismus guidelines and Alaska Blind Child Discovery institutional review board protocol. Instrument referral guidelines determined through phase 1 comparison was then applied on additional patients for validation in phase 2. RESULTS: Sixty-two children (age: 1 to 10 years, mean: 5.2 years) were screened with all three devices before cycloplegic examination. Refractive results were inconclusive due to pupil size, cooperation, and out-of-range values. Values for sensitivity (91% and 78%), specificity (71% and 59%), and inconclusive rate (10% and 13%) were found for PlusoptiX and SPOT. The 2WIN was calibrated for this age range (phase 1), yielding 71% sensitivity, 67% specificity, and a 5% inconclusive rate. Regression compared to examination for the PlusoptiX, SPOT, and 2WIN, respectively, were sphere (r(2): 0.76, 0.87, and 0.58), cylinder power (r(2): 0.67, 0.56, and 0.50), and cylinder axis (r(2): 0.71, 0.41, and 0.40). A preferred 2WIN instrument criteria set was determined from the receiver operating characteristic curve. In phase 2, with 117 patients comparing 2WIN to PlusoptiX A-09, sensitivity was 73% and 85%, specificity was 76% and 73%, and inconclusive rate was 8% and 12%, respectively. The three instant-interpreting photorefractors performed well on amblyopic children, with the 2WIN having low inconclusive results. The PlusoptiX outperformed the SPOT and 2WIN as an autorefractor, particularly with respect to astigmatism power and axis. CONCLUSIONS: The new 2WIN is a promising addition to portable photoscreeners for amblyopia detection and estimating refractive error.