IS711-based real-time PCR assay as a tool for detection of Brucella spp. in wild boars and comparison with bacterial isolation and serology

<p>Abstract</p> <p>Background</p> <p>Control of brucellosis in livestock, wildlife and humans depends on the reliability of the methods used for detection and identification of bacteria. In the present study, we describe the evaluation of the recently established real-time PCR assay based on the <it>Brucella</it>-specific insertion sequence IS<it>711 </it>with blood samples from 199 wild boars (first group of animals) and tissue samples from 53 wild boars (second group of animals) collected in Switzerland. Results from IS<it>711 </it>real-time PCR were compared to those obtained by bacterial isolation, Rose Bengal Test (RBT), competitive ELISA (c-ELISA) and indirect ELISA (i-ELISA).</p> <p>Results</p> <p>In the first group of animals, IS<it>711 </it>real-time PCR detected infection in 11.1% (16/144) of wild boars that were serologically negative. Serological tests showed different sensitivities [RBT 15.6%, c-ELISA 7.5% and i-ELISA 5.5%] and only 2% of blood samples were positive with all three tests, which makes interpretation of the serological results very difficult. Regarding the second group of animals, the IS<it>711 </it>real-time PCR detected infection in 26% of animals, while <it>Brucella </it>spp. could be isolated from tissues of only 9.4% of the animals.</p> <p>Conclusion</p> <p>The results presented here indicate that IS<it>711 </it>real-time PCR assay is a specific and sensitive tool for detection of <it>Brucella </it>spp. infections in wild boars. For this reason, we propose the employment of IS<it>711 </it>real-time PCR as a complementary tool in brucellosis screening programs and for confirmation of diagnosis in doubtful cases.</p

A Case Study of Zoonotic <i>Chlamydia abortus</i> Infection: Diagnostic Challenges From Clinical and Microbiological Perspectives

Chlamydia abortus is the most common causative agent of abortion in small ruminants, but it is poorly recognized as a human pathogen. In most published case studies, diagnosis remained difficult and often resulted in delayed initiation of therapy. In this case study of severe C abortus infection in a pregnant farmer from Switzerland, we highlight the clinical and microbiological diagnostic challenges and provide evidence of a zoonotic epidemiological link

A case report of a cystic fibrosis patient with repeated isolation of Trichosporon mycotoxinivorans identified by a novel short-extraction method

Abstract Background Trichosporon mycotoxinivorans is a recently described yeast-like fungal organism and its association as a pathogen for patients with cystic fibrosis (CF) was reported previously. We show the clinical course of a CF patient over 9 years as well as the applications of modern molecular and proteomic identification techniques of this rare fungus. Case presentation We present the case of a 32-year-old male CF patient with sputum cultures continuously positive with the anamorphic yeast T. mycotoxinivorans during 9 years. Furthermore, susceptibility testing of T. mycotoxinivorans to different antifungals were performed. In addition, a rapid identification method of this novel fungal pathogen with matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) was applied using a simple extraction protocol. Conclusions Our case presentation confirms T. mycotoxinivorans as a potential emerging pathogen in patients with CF. However, our CF patient showed mild symptoms over a very long time period of 9 years. A short MALDI-TOF MS procedure allows reliable and rapid identification of T. mycotoxinivorans and therefore should facilitate further study on the clinical relevance and epidemiology of this unusual fungal organism

Ultrasensitive Detection of Mutations and Genes Relevant to Antimicrobial Resistance in Bacteria

The worldwide emergence of multidrug‐resistant (MDR) bacteria is associated with significant morbidity, mortality, and healthcare costs. Rapid and accurate diagnostic methods to detect antibiotic resistance are critical for antibiotic stewardship and infection control measurements. Here a cantilever nanosensor‐based diagnostic assay is shown to detect single nucleotide polymorphisms (SNPs) and genes associated with antibiotic resistance in Gram negative ( Pseudomonas aeruginosa ) and positive ( Enterococcus faecium ) bacteria, representing frequent causes for MDR infections. Highly specific RNA capture probes for SNPs ( ampR D135G or ampR G154R ) or resistance genes ( vanA , vanB , and vanD ) allow to detect the binding of bacterial RNA within less than 5 min. Serial dilutions of bacterial RNA indicate an unprecedented sensitivity of 10 fg µL −1 total RNA corresponding to less than ten bacterial cells for SNPs and 1 fg µL −1 total RNA for vanD detection equivalent to single bacterial cell sensitivity