False positive molecular diagnosis of Leber's hereditary optic neuropathy

M. Y. Yen, A. G. Wang, W. L. Chang, Wen-Ming Hsu, J. H. Liu, Y. H. Wei

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Background. The most common pathogenic mitochondrial DNA (mtDNA) mutation associated with Leber's hereditary optic neuropathy (LHON) is at the 11,778 nucleotide (nt) position and is usually detected by loss of an Sfa NI restriction site. However, Sfa NI restriction site includes five nucleotides. Substitution of any of the five nucleotides leads to loss of the cutting site and causes a false-positive result. We investigated the false-positive diagnosis of LHON by loss of the Sfa NI restriction site using Sfa NI restriction site analysis and single-strand conformation polymorphism (SSCP) analysis. Methods. Mae III restriction analysis for double confirmation of the Sfa NI restriction site and direct sequencing for final confirmation of SSCP analysis were performed. Results. The sensitivity of Sfa NI test was 100% and the specificity of the Sfa NI test was 97%. The false-positive rate of Sfa NI test was 3%. SSCP analysis showed 100% sensitivity. Direct sequencing showed 32 patients had a mutation at nt 11,778 of mtDNA and one patient had a silent mutation at nt 11,782 of mtDNA. Conclusions. These results suggest that restriction enzyme digestion analysis requires double confirmation to avoid a false-positive diagnosis and that DNA sequencing is needed for the confirmation of the mutation detected by SSCP.

Original languageEnglish
Pages (from-to)864-868
Number of pages5
JournalChinese Medical Journal (Taipei)
Volume63
Issue number12
Publication statusPublished - Dec 1 2000
Externally publishedYes

Fingerprint

Leber's Hereditary Optic Atrophy
Nucleotides
Mitochondrial DNA
Mutation
Restriction Mapping
DNA Sequence Analysis
Digestion

Keywords

  • Mitochondrial DNA
  • Optic atrophy
  • Point mutation
  • Restriction mapping

ASJC Scopus subject areas

  • Medicine(all)

Cite this

Yen, M. Y., Wang, A. G., Chang, W. L., Hsu, W-M., Liu, J. H., & Wei, Y. H. (2000). False positive molecular diagnosis of Leber's hereditary optic neuropathy. Chinese Medical Journal (Taipei), 63(12), 864-868.

False positive molecular diagnosis of Leber's hereditary optic neuropathy. / Yen, M. Y.; Wang, A. G.; Chang, W. L.; Hsu, Wen-Ming; Liu, J. H.; Wei, Y. H.

In: Chinese Medical Journal (Taipei), Vol. 63, No. 12, 01.12.2000, p. 864-868.

Research output: Contribution to journalArticle

Yen, MY, Wang, AG, Chang, WL, Hsu, W-M, Liu, JH & Wei, YH 2000, 'False positive molecular diagnosis of Leber's hereditary optic neuropathy', Chinese Medical Journal (Taipei), vol. 63, no. 12, pp. 864-868.
Yen, M. Y. ; Wang, A. G. ; Chang, W. L. ; Hsu, Wen-Ming ; Liu, J. H. ; Wei, Y. H. / False positive molecular diagnosis of Leber's hereditary optic neuropathy. In: Chinese Medical Journal (Taipei). 2000 ; Vol. 63, No. 12. pp. 864-868.
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abstract = "Background. The most common pathogenic mitochondrial DNA (mtDNA) mutation associated with Leber's hereditary optic neuropathy (LHON) is at the 11,778 nucleotide (nt) position and is usually detected by loss of an Sfa NI restriction site. However, Sfa NI restriction site includes five nucleotides. Substitution of any of the five nucleotides leads to loss of the cutting site and causes a false-positive result. We investigated the false-positive diagnosis of LHON by loss of the Sfa NI restriction site using Sfa NI restriction site analysis and single-strand conformation polymorphism (SSCP) analysis. Methods. Mae III restriction analysis for double confirmation of the Sfa NI restriction site and direct sequencing for final confirmation of SSCP analysis were performed. Results. The sensitivity of Sfa NI test was 100{\%} and the specificity of the Sfa NI test was 97{\%}. The false-positive rate of Sfa NI test was 3{\%}. SSCP analysis showed 100{\%} sensitivity. Direct sequencing showed 32 patients had a mutation at nt 11,778 of mtDNA and one patient had a silent mutation at nt 11,782 of mtDNA. Conclusions. These results suggest that restriction enzyme digestion analysis requires double confirmation to avoid a false-positive diagnosis and that DNA sequencing is needed for the confirmation of the mutation detected by SSCP.",
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AU - Yen, M. Y.

AU - Wang, A. G.

AU - Chang, W. L.

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AU - Liu, J. H.

AU - Wei, Y. H.

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N2 - Background. The most common pathogenic mitochondrial DNA (mtDNA) mutation associated with Leber's hereditary optic neuropathy (LHON) is at the 11,778 nucleotide (nt) position and is usually detected by loss of an Sfa NI restriction site. However, Sfa NI restriction site includes five nucleotides. Substitution of any of the five nucleotides leads to loss of the cutting site and causes a false-positive result. We investigated the false-positive diagnosis of LHON by loss of the Sfa NI restriction site using Sfa NI restriction site analysis and single-strand conformation polymorphism (SSCP) analysis. Methods. Mae III restriction analysis for double confirmation of the Sfa NI restriction site and direct sequencing for final confirmation of SSCP analysis were performed. Results. The sensitivity of Sfa NI test was 100% and the specificity of the Sfa NI test was 97%. The false-positive rate of Sfa NI test was 3%. SSCP analysis showed 100% sensitivity. Direct sequencing showed 32 patients had a mutation at nt 11,778 of mtDNA and one patient had a silent mutation at nt 11,782 of mtDNA. Conclusions. These results suggest that restriction enzyme digestion analysis requires double confirmation to avoid a false-positive diagnosis and that DNA sequencing is needed for the confirmation of the mutation detected by SSCP.

AB - Background. The most common pathogenic mitochondrial DNA (mtDNA) mutation associated with Leber's hereditary optic neuropathy (LHON) is at the 11,778 nucleotide (nt) position and is usually detected by loss of an Sfa NI restriction site. However, Sfa NI restriction site includes five nucleotides. Substitution of any of the five nucleotides leads to loss of the cutting site and causes a false-positive result. We investigated the false-positive diagnosis of LHON by loss of the Sfa NI restriction site using Sfa NI restriction site analysis and single-strand conformation polymorphism (SSCP) analysis. Methods. Mae III restriction analysis for double confirmation of the Sfa NI restriction site and direct sequencing for final confirmation of SSCP analysis were performed. Results. The sensitivity of Sfa NI test was 100% and the specificity of the Sfa NI test was 97%. The false-positive rate of Sfa NI test was 3%. SSCP analysis showed 100% sensitivity. Direct sequencing showed 32 patients had a mutation at nt 11,778 of mtDNA and one patient had a silent mutation at nt 11,782 of mtDNA. Conclusions. These results suggest that restriction enzyme digestion analysis requires double confirmation to avoid a false-positive diagnosis and that DNA sequencing is needed for the confirmation of the mutation detected by SSCP.

KW - Mitochondrial DNA

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