Table Study profiles of included studies on diagnostic accuracy Inclusion criteria / Study Study design Study population Index test(s) Reference standard Exclusion criteria Country Quality Axton & Level IV: N=193 mouthwash samples None reported Restriction generation PCR. This technique No reference standard Brock () from CF patients uses a mismatch primer to introduce a base A study of diagnostic substitution designed to create or destroy a UK yield (no reference restriction site when a mutation is present. standard) Quality: poor with a high risk of bias Bareil et al. () Level III-2: N=182 samples from men Inclusion criteria: A complete scan of the 27 coding/flanking Clinical diagnosis of with a clinical diagnosis of sequences of the CFTR gene was performed CBAVD was based on France A comparison with Clinical diagnosis of CBAVD CBAVD by either DGGE or DHPLC. clinical examination with reference standard (not Exclusion criteria: impalpable vas deferens, blinded or blinding not In addition, two intronic mutations, Patients with renal transrectal ultrasonography, known) 18111.6kbAG in IVS11 and 384910kbCT in abnormalities semen analysis and sperm IVS19, and variations at locus IVS8-Tn were Quality: poor with a high count in accordance with screened by specific PCR restriction tests. risk of bias World Health Organization Samples with only 1 or no CFTR guidelines. mutations were further investigated for large rearrangements by a semi-quantitative fluorescent PCR assay. Samples showing abnormal profiles were directly sequenced with BigDye Terminator sequencing. Bernardino, Level III-2: N=17 patients with CBAVD Inclusion criteria: Mutation detection studies were carried out on Diagnosis of CBAVD was Lima & Zatz all 27 exons and exon–intron boundaries of the based on scrotal A comparison with 20 unrelated men (ages () CFTR gene by SSCP and heteroduplex examination, ultrasound reference standard (not 49 years) with obstructive analysis. and semen analysis. Brazil blinded or blinding not azoospermia; 17 were known) diagnosed with CBAVD; none CFTR gene variants of the T tract length of had been diagnosed with CF intron 8 were also investigated. Quality: poor with a high risk of bias Abnormally migrating fragments were subsequently sequenced to confirm the
Table Study profiles of included studies on diagnostic accuracy Study Country Study design Quality Study population Inclusion criteria / Exclusion criteria Index test(s) Reference standard Axton & Brock () UK Level IV: A study of diagnostic yield (no reference standard) Quality: poor with a high risk of bias N=193 mouthwash samples from CF patients None reported Restriction generation PCR. This technique uses a mismatch primer to introduce a base substitution designed to create or destroy a restriction site when a mutation is present. No reference standard Bareil et al. () France Level III-2: A comparison with reference standard (not blinded or blinding not known) Quality: poor with a high risk of bias N=182 samples from men with a clinical diagnosis of CBAVD Inclusion criteria: Clinical diagnosis of CBAVD Exclusion criteria: Patients with renal abnormalities A complete scan of the 27 coding/flanking sequences of the CFTR gene was performed by either DGGE or DHPLC. In addition, two intronic mutations, 18111.6kbAG in IVS11 and 384910kbCT in IVS19, and variations at locus IVS8-Tn were screened by specific PCR restriction tests. Samples with only 1 or no CFTR mutations were further investigated for large rearrangements by a semi-quantitative fluorescent PCR assay. Samples showing abnormal profiles were directly sequenced with BigDye Terminator sequencing. Clinical diagnosis of CBAVD was based on clinical examination with impalpable vas deferens, transrectal ultrasonography, semen analysis and sperm count in accordance with World Health Organization guidelines. Bernardino, Lima & Zatz () Brazil Level III-2: A comparison with reference standard (not blinded or blinding not known) Quality: poor with a high risk of bias N=17 patients with CBAVD Inclusion criteria: 20 unrelated men (ages years) with obstructive azoospermia; 17 were diagnosed with CBAVD; none had been diagnosed with CF Mutation detection studies were carried out on all 27 exons and exon–intron boundaries of the CFTR gene by SSCP and heteroduplex analysis. CFTR gene variants of the T tract length of intron 8 were also investigated. Abnormally migrating fragments were subsequently sequenced to confirm the Diagnosis of CBAVD was based on scrotal examination, ultrasound and semen analysis. 49
1216-MSAC-CA-CFTR.docx