Rett Syndrome: MECP2 Targeted Analysis

Clinical Information

A neurodevelopmental disorder that affects females, Rett syndrome is associated with cortical atrophy, stereotypical hand movements and severe intellectual disability. With an incidence of 1 in 10,000 – 15,000, it is one of the most common causes of intellectual disability in females. Rett syndrome is characterized by loss of acquired skills after a period of normal development in infancy.

Most cases of Rett syndrome are caused by an abnormality (missense variant, nonsense variant or deletion) of MECP2. Additionally, a specific phenotype has been identified in males with a MECP2 duplication that is identifiable by MLPA. These males display hypotonia that progresses to spasticity, severe intellectual disability and recurrent pulmonary infections. Females in these families who have the duplication are clinically unaffected and display a near total skewing of X-inactivation. Sequencing of the MECP2 gene and MLPA deletion/duplication analysis can be ordered concurrently, sequentially or separately.

Technical Information

• This test is performed by Sanger sequencing.
• Genomic DNA is used to amplify the region of interest of the MECP2 gene by standard polymerase chain reaction. The amplified product is sequenced using the Sanger dideoxy method.

Specimen Requirements

• The preferred sample type is 3-4 ml of peripheral blood collected in an EDTA (purple top) tube. Dried blood spots, extracted DNA, and saliva are also accepted for this test. Blood and saliva kits are available by request.
• Approximately 75 µL of blood should be applied to each of the five circles on a filter paper dried blood spot card. Allow blood to dry for at least 4 hours before shipping.
• Send approximately 5 µg of extracted DNA at a requested concentration of 90-130 ng/µl.
• Saliva samples must be submitted in an approved saliva kit.

Transport Instructions

• The blood specimen should be kept at room temperature and delivered via overnight shipping. If shipment is delayed by one or two days, the specimen should be refrigerated and shipped at room temperature. Do not freeze the specimen. Samples collected on Friday can be safely designated for Monday delivery.
• DBS cards should be shipped at room temperature.
• Extracted DNA should be sent at room temperature via overnight delivery.
• Saliva is stable at room temperature and can be delivered via overnight or ground shipping.

Prenatal Testing Information

Prenatal diagnosis may be available for familial pathogenic or likely pathogenic variants; full sequencing of the MECP2 gene is not available. For variants identified at external laboratories, GGC will need to review the variant classification prior to sample receipt. Prenatal diagnosis is not available for variants of uncertain clinical significance. Additional fees for cell culture and maternal cell contamination may apply. Maternal cell contamination studies are required for all prenatal molecular tests. Contact the laboratory prior to sending a prenatal specimen to confirm that targeted testing can be accepted.

Prenatal Specimen Requirements

• Amniotic Fluid: Direct amniotic fluid will be accepted for analysis given there is sufficient volume for back-up culture to be established. A back-up culture at a reference lab OR Greenwood lab is required. Additional charges may apply if cell culture is required after receipt of the sample. If sending direct fluid for molecular analysis only, 10-20 ml of amniotic fluid is requested. Chromosome studies will require an additional 10-15 ml of fluid.
• Chorionic Villi Sample (CVS): 10-50 mg of chorionic villi is requested in a sterile tube with CVS media. Direct testing on CVS is not available; an additional 1-3 weeks may be needed for cultures to grow.
• Cultured amniocytes/CVS: 2x T25 confluent flasks
• A maternal sample is required, and accepted sample types include blood, saliva, and extracted DNA. Please refer to the specimen requirements above for more information.

Prenatal Transport Instructions

• Amniotic fluid should be kept at room temperature; do not freeze or refrigerate. Specimen should be sent by courier or overnight mail to arrive at the laboratory the next day. FedEx is preferred.
• CVS and cultured amniocytes should be kept at room temperature; do not freeze or refrigerate. Specimen should be sent by courier or overnight mail to arrive at the laboratory the next day. FedEx is preferred.
• Please refer to the transport instructions above for sending the maternal sample.