| 1. |
- Hahn, Katharina, et al.
(author)
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Establishing and validating the fluorescent amyloid ligand h-FTAA (heptamer formyl thiophene acetic acid) to identify transthyretin amyloid deposits in carpal tunnel syndrome
- 2017
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In: Amyloid. - : TAYLOR & FRANCIS LTD. - 1350-6129 .- 1744-2818. ; 24:2, s. 78-86
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Journal article (peer-reviewed)abstract
- Transthyretin-derived (ATTR) amyloidosis is a frequent finding in carpal tunnel syndrome. We tested the following hypotheses: the novel fluorescent amyloid ligand heptameric formic thiophene acetic acid (h-FTAA) has a superior sensitivity for the detection of amyloid compared with Congo red-staining; Amyloid load correlates with patient gender and/or patient age. We retrieved 208 resection specimens obtained from 184 patients with ATTR amyloid in the carpal tunnel. Serial sections were stained with Congo red, h-FTAA and an antibody directed against transthyretin (TTR). Stained sections were digitalized and forwarded to computational analyses. The amount of amyloid was correlated with patient demographics. Amyloid stained intensely with h-FTAA and an anti-TTR-antibody. Congo red-staining combined with fluorescence microscopy was significantly less sensitive than h-FTAA-fluorescence and TTR-immunostaining: the highest percentage area was found in TTR-immunostained sections, followed by h-FTAA and Congo red. The Pearson correlation coefficient was .8 (Congo red vs. h-FTAA) and .9 (TTR vs. h-FTAA). Amyloid load correlated with patient gender, anatomical site and patient age. h-FTAA is a highly sensitive method to detect even small amounts of ATTR amyloid in the carpal tunnel. The staining protocol is easy and h-FTAA may be a much more sensitive procedure to detect amyloid at an earlier stage.
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| 2. |
- Kieninger, Barbara, et al.
(author)
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PTAA and B10 : new approaches to amyloid detection in tissue-evaluation of amyloid detection in tissue with a conjugated polyelectrolyte and a fibril-specific antibody fragment
- 2011
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In: Amyloid. - : Informa UK Limited. - 1350-6129 .- 1744-2818. ; 18:2, s. 47-52
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Journal article (peer-reviewed)abstract
- Methods: aEuro integral We compared the amyloid detection of PTAA and B10 to Congo red in 106 amyloid-containing tissue biopsies of diverse anatomical and precursor origin by evaluating the accordance in four grades (grade 0: no staining, grade 1: staining of < 33%% of the amyloid deposits, grade 2: 33--66%% and grade 3: aEuroS > 66%%). Results: aEuro integral PTAA showed grade 2--3 staining in 57 (54%%) cases, while B10 presented this accordance in only 25 (24%%) tissue biopsies. Grade 1 staining was found in 11 (10%%) samples with PTAA and in 62 (58%%) cases with B10. No staining at all (grade 0) occurred in 38 (36%%) biopsies when using PTAA and in 19 (18%%) cases when using B10. Conclusion: aEuro integral Although conformation-sensitive detection seemed promising, PTAA and B10 stain only a fraction of the examined amyloid samples when using routine surgical pathology settings. This study emphasises the necessity of having optimised pre-analytical protocols for recovery, storage and handling of samples if these novel amyloid ligands are to be used in routine diagnosis of amyloid.
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| 3. |
- Sjölander, Daniel, et al.
(author)
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Establishing the fluorescent amyloid ligand h-FTAA for studying human tissues with systemic and localized amyloid
- 2016
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In: Amyloid. - : Informa UK Limited. - 1350-6129 .- 1744-2818. ; 23:2, s. 98-108
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Journal article (peer-reviewed)abstract
- Rapid and accurate detection of amyloid deposits in routine surgical pathology settings are of great importance. The use of fluorescence microscopy in combination with appropriate amyloid specific dyes is very promising in this regard. Here we report that a luminescent conjugated oligothiophene, h-FTAA, rapidly and with high sensitivity and selectivity detects amyloid deposits in verified clinical samples from systemic amyloidosis patients with AA, AL and ATTR types; as well as in tissues laden with localized amyloidosis of AANF, AIAPP and ASem1 type. The probe h-FTAA emitted yellow red fluorescence on binding to amyloid deposits, whereas no apparent staining was observed in surrounding tissue. The only functional structure stained with h-FTAA showing the amyloidotypic fluorescence spectrum was Paneth cell granules in intestine. Screening of 114 amyloid containing tissues derived from 107 verified (Congo red birefringence and/or immunohistochemistry) amyloidosis patients revealed complete correlation between h-FTAA and Congo red fluorescence (107/107, 100% sensitivity). The majority of Congo red negative control cases (27 of 32, 85% specificity) were negative with h-FTAA. Small Congo red negative aggregates in kidney, liver, pancreas and duodenum were found by h-FTAA fluorescence in five control patients aged 72-83 years suffering from diverse diseases. The clinical significance of these false-positive lesions is currently not known. Because h-FTAA fluorescence is one magnitude brighter than Congo red and as the staining is performed four magnitudes lower than the concentration of dye, we believe that these inclusions are beyond detection by Congo red. We conclude that h-FTAA is a fluorescent hypersensitive, rapid and powerful tool for identifying amyloid deposits in tissue sections. Use of h-FTAA can be exploited as a rapid complementary technique for accurate detection of amyloid in routine surgical pathology settings. Our results also implicate the potential of the technique for detection of prodromal amyloidosis as well as for discovery of new amyloid-like protein aggregates in humans.
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