Strom et al. Level III-2 Carrier screening CF testing for the ACMG Irregularities such as unusually weak None reported There were 9 instances of 2003 program: number of panel using the Roche CF lines or missing lines can be present on ambiguous results (approximately 1 Low risk of patients screened not Gold Linear Array strips the strip assay, and these ambiguities per 10,000 assays) USA bias reported can be resolved by sequencing the appropriate amplicon. Giuliani et Level III-2 N=23 CBAVD patients MLPA to detect deletions Large deletions involving the CFTR MLPA detected 2 deletion Not reported al. (2010) and/or duplications of the gene are not detectable by any mutations in 2 CBAVD Some risk of CFTR gene sequencing assay. patients that could not be Italy bias detected with DHPLC plus DHPLC with abnormal sequencing. results were analysed by direct DNA sequencing to detect point mutations of the CFTR gene. Houdayer et Level III-2 N=40 DNA samples of CF(12) ARMS kit ARMS technology cannot distinguish 2/40 patients with Not reported known CFTR between heterozygous and homozygous mutations were al. (1998) Some risk of mutations homozygous point mutations. not identified as homozygous. France bias Level III-2 N=133 DNA samples SSCP analysis of PCR The mobility shift of single strands is There was no optimal Not reported et al. of known CFTR products dependent on electrophoretic condition suitable for all High risk of (1994) mutations conditions such as temperature, ionic exons. However, using a 10% bias strength, composition of the gel, gel with 1.3% cross-linking in USA addition of glycerol. the presence of glycerol had a sensitivity of 100% for most The sensitivity of SSCP varied greatly exons and 80–98% for exons with the conditions and between exons. 4, 7 and 13. Level III-2 N=73 DNA samples of DHPLC using Stanford An increase of the denaturation 7/73 (9.6%) mutations not Not reported et al. known CFTR MELT software temperature by 1–2 degrees above that detected at the recommended High risk of (2002) mutations recommended by the MELT program denaturation temperature bias improved the resolution of mutation USA detection to 100%.
Strom et al. 2003 USA Level III-2 Low risk of bias Carrier screening program: number of patients screened not reported CF testing for the ACMG panel using the Roche CF Gold Linear Array strips Irregularities such as unusually weak lines or missing lines can be present on the strip assay, and these ambiguities can be resolved by sequencing the appropriate amplicon. None reported There were 9 instances of ambiguous results (approximately 1 per 10,000 assays) Giuliani et al. (2010) Italy Level III-2 Some risk of bias N=23 CBAVD patients MLPA to detect deletions and/or duplications of the CFTR gene DHPLC with abnormal results were analysed by direct DNA sequencing to detect point mutations of the CFTR gene. Large deletions involving the CFTR gene are not detectable by any sequencing assay. MLPA detected 2 deletion mutations in 2 CBAVD patients that could not be detected with DHPLC plus sequencing. Not reported Houdayer et al. (1998) France Level III-2 Some risk of bias N=40 DNA samples of known CFTR mutations CF(12) ARMS kit ARMS technology cannot distinguish between heterozygous and homozygous point mutations. 2/40 patients with homozygous mutations were not identified as homozygous. Not reported et al. (1994) USA Level III-2 High risk of bias N=133 DNA samples of known CFTR mutations SSCP analysis of PCR products The mobility shift of single strands is dependent on electrophoretic conditions such as temperature, ionic strength, composition of the gel, addition of glycerol. The sensitivity of SSCP varied greatly with the conditions and between exons. There was no optimal condition suitable for all exons. However, using a 10% gel with 1.3% cross-linking in the presence of glycerol had a sensitivity of 100% for most exons and 80–98% for exons 4, 7 and 13. Not reported et al. (2002) USA Level III-2 High risk of bias N=73 DNA samples of known CFTR mutations DHPLC using Stanford MELT software An increase of the denaturation temperature by 1–2 degrees above that recommended by the MELT program improved the resolution of mutation detection to 100%. 7/73 (9.6%) mutations not detected at the recommended denaturation temperature Not reported