NEI DNA Diagnostics Laboratory Operation CLIA recertification: The most important clinical activity in NEI DNA Diagnostic Laboratory in 2009 is the successful recertification through CMS regional CLIA office. NEI DNA lab suspended DNA diagnostic testing activity since 2007 and only maintained the eyeGENE core facility function for sample processing because of the directorship situation. With the new leadership, NEI DNA lab re-built the genetic testing component in early 2009 and resumed clinical testing in may 2009 through a newly established standard QA (Quality Assurance) procedure in a compliance with the CLIA law and regulations. This license was expired in November, 2009 and subject to be renewed. To prepare for the inspection for recertification, NEI DNA lab reviewed, revised, and redesigned entire protocol menu. The lab was successfully inspected on Dec 2, 2009. NEI DNA lab is now recertified for another two years. NEI DNA lab facilities: To comply with the CLIA regulations for facility requirement, with the great supports from the NEI Intramural Scientific Director and Deputy Scientific Director, the laboratory was eventually relocated and expanded to current assignments in 2009. It is critical for the lab to operate four different operations, eyeGENE, NEI DNA lab, resequencing project and lab research with current space, equipments, and supplies because of the potential difficulties and risks otherwise. Revalidation: During the reinstallation of genetic testing, the new testing biologist was sent to Applied Biosystems Inc training class at Rockville for 3130xl sequencer use. He was trained for clinical procedures and managing of equipments for CLIA purposes. And he was also trained to serve as lab manager to hold lab credit card for purchasing purposes. The first re-validated protocol was the DNA testing for the gene responsible for the X-linked Juvenile Retinoschisis, XLRS1 gene. This re-validation was completed in May, 2009. Since then, the lab re-validated testing for CYP4V2, gene responsible for Bietti Crystalline Corneoretinal Dystrophy. Re-validation of another gene, RPGR, responsible for X-linked Retinitis Pigmentosa, RP3 is ongoing. Validation of NYX (Nyctalopin) testing is about ready to offer. The job of re-validation of RPGR and NYX took a lot of time. Especially, one of the major target exon, ORF15, is 1.7 kb long. Many publications demonstrated and other CLIA labs in eyeGENE network experienced the technical difficulties in PCR amplifications of this exon. Now the lab is making progress and about to finish the re-validation of NYX. We will move on finishing up re-validation of RPGR. Resolving backlog: There was a backlog from the past years. Many submitted samples were waiting for re-installation of DNA testing. After re-validation of XLRS1, the lab has completed 21 clinical reports including reports to eyeGENE program in 2009 and 29 reports (up to now) in 2010. The lab has completed CYP4V2 gene testing for seven patients since the revalidation. Developing new clinical tests: NEI DNA lab has development seven new additional clinical protocols for sample analysis. The lab commits to continue the development of new clinical tests. Now the lab is working on a gene responsible for Renal-coloboma syndrome, PAX2 gene with help from new OGVFB fellow. The lab is planning to develop several gene tests, including CHM, EYS, and PSTAN12. Inventory of DNA storage: The lab identified earlier that the eyeGENE program should have a more efficient procedure to organize and track any changes in DNA storage. The eyeGENE program has just reached 2000 samples milestone. It would be very important to be more efficient and organized. Another round of laboratory inventory was completed in the summer 2010. NEI DNA laboratory storage and tracking system is more reliable. The lab is granted with a new automated DNA extraction processor. A new protocol based on the equipment is under development. Sample switch investigation: Last September NEI DNA lab identified an incident of sample switch involving both eyeGENE samples and NEI DNA lab samples. A female DNA sample was sent to another CLIA lab for referred testing under the name of a male patient. After the incident, the sample was recalled and fresh blood from the male patient was sent to the reference lab for the testing. Meanwhile, fresh blood from the male patient was requested again for a thorough quality assessment investigation by comparing with recalled samples and stored blood. During the investigation, NEI DNA lab had to develop a series of new testing protocols for the required analyses for the investigation first. We developed a direct PCR from limited blood assay, three STR markers on X chromosome assays, and three autosomal chromosome STR markers assay for identity analysis. We have analyzed hundreds of results. During the investigation, two eyeGENE samples were found to be switched each other in one of the two sets of DNA storage. Two NEI banking samples were found having same ID but were identified as from different individuals. A female patient blood was received in NEI DNA lab, but blood or DNA can not be located. The lab has been in the process to replace storage with new requested and confirmed DNA samples. The lab commits to zero tolerance for any sample processing uncertainty and will conduct very thorough investigation for every incidence and replace every sample in question.
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