Cervical dystonia and no oculomotor apraxia as new manifestation of ataxia-telangiectasia-like disorder 1 – case report and review of the literature

Ataxia-telangiectasia-like disorder 1 (ATLD1) is a rare neurodegenerative disorder associated with early onset ataxia and oculomotor apraxia. The genetic determination of ATLD1 is a mutation in the MRE11 gene (meiotic recombination 11 gene), which causes DNA-double strand break repair deficits. Clinical features of patients with ATLD1 resemble those of ataxia telangiectasia (AT), with slower progression and milder presentation. Main symptoms include progressive cerebellar ataxia, oculomotor apraxia, cellular hypersensitivity to ionizing radiations. Facial dyskinesia, dystonia, dysarthria have also been reported. Here we present a 45-year old woman with cervical and facial dystonia, dysarthria and ataxia, who turned out to be the first case of ATLD without oculomotor apraxia, and with dystonia as a main manifestation of the disease. She had presented those non-specific symptoms for years, before whole exome sequencing confirmed the diagnosis

Development of a Sensitive and Specific Polyclonal Antibody for Serological Detection of Clavibacter michiganensis subsp. sepedonicus.

The quarantine bacterium Clavibacter michiganensis subsp. sepedonicus (Cms) causes bacterial ring rot (BRR) in potato but is difficult to detect, hampering the diagnosis of this disease. ELISA immunoassays have not been widely used to detect Cms because commercially available anti-Cms antibodies detect mainly EPS-producing bacteria and can fail to detect strains that do not produce EPS. In the current study, we developed a new type of polyclonal antibody that specifically detects Clavibacter michiganensis subsp. sepedonicus bacteria irrespective of their EPS level. We first found that the presence of bacterial EPS precluded quantitative measurement of bacteria by currently available immunoenzymatic methods, but that washing Cms cells with acidic and basic buffers to remove EPS before analysis successfully standardized ELISA results. We used a mix of three strains of Cms with diverse EPS levels to generate antigen for production of antibodies recognizing Cms cells with and without an EPS layer (IgG-EPS and IgG-N-EPS, respectively). The resulting IgG-N-EPS recognized almost all Cms strains tested in this work regardless of their mucoidal level. The availability of this new antibody renders immunological diagnostics of Cms more sensitive and reliable, as our newly developed antibodies can be used in many type of immunoassays. This work represents an important step forward in efforts to diagnose and prevent the spread of BRR, and the methods and solutions developed in this work are covered by six Polish, one European and one US patents

Detection of <i>Clavibacter michiganensis</i> subsp. <i>sepedonicus</i> mucoid and nonmucoid cells by PTA-ELISA with the newly developed antibodies.

<p>Assays were carried out with different titers of the obtained IgG against mucoid and nonmucoid cells. Immunization mixtures of strains NCPPB 4053, 758 and 527 were used at 10⁵ CFU/ml.</p

Colony morphology of the <i>Clavibacter michiganensis</i> subsp. <i>sepedonicus</i> strains selected for production of polyclonal antibodies in rabbit, according to their EPS levels.

<p>Left. Strain 527 –Rough colony type. Middle. Strain 758 –Intermediate colony type. Right. Strain NCPPB 4053 –Fluidal colony type.</p

Detection of different Cms strains in terms of their EPS levels by DAS-ELISA (Loewe kit).

<p>Bacterial cells of three different strains (527 –Rough, 758 –Intermediate, NCPPB 4053 –Fluidal) were suspended: (1) in water, (2) washed three times with ddH₂O, (3) washed twice at pH = 2.5 and three times with ddH₂O, (4) washed twice at pH = 10.5 and three times with ddH₂O, (5) washed twice at pH = 2.5, twice at pH = 10.5, and three times with ddH₂O.Absorbance equal to at least two-fold the value of the blank sample was accepted as a positive result. All assays were performed twice in three replicates.</p

Comparison of the sensitivity of different ELISA tests for detecting Cms bacterial cells.

<p>Strains tested were 527 and NCPPB 4053.</p