Genotyping ofBorreliafrom formalin‐fixed paraffin‐embedded skin biopsies of cutaneous borreliosis and tick bite reactions by assays targeting the intergenic spacer region,ospAandospCgenes

Brandt, F.C.; Ertas, Beyhan; Falk, Thomas; Metze, Dieter; Böer‐Auer, Almut

doi:10.1111/bjd.12855

Cited by 18 publications

(25 citation statements)

References 51 publications

(104 reference statements)

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“…In humans, the most common manifestation of Lyme borreliosis is erythema migrans (39). This skinassociated lesion is often linked to the presence of B. afzelii (8,40,41), the most common species identified in patient samples in our institute (A. Schötta, M. Reiter, I. Korschinek, A. Müller, G. Khanakah, H. Stockinger, and G. Stanek, presented at the 33rd Annual Meeting of the Austrian Society for Hygiene, Microbiology and Preventive Medicine, Salzburg, Austria, 21 to 24 May 2012).…”

Section: Discussionmentioning

confidence: 99%

Approaches for Reverse Line Blot-Based Detection of Microbial Pathogens in Ixodes ricinus Ticks Collected in Austria and Impact of the Chosen Method

Schötta

Wijnveld

Stockinger

et al. 2017

Appl Environ Microbiol

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Ticks transmit a large number of pathogens capable of causing human disease. In this study, the PCR-reverse line blot (RLB) method was used to screen for pathogens in a total of 554 Ixodes ricinus ticks collected from all provinces of Austria. These pathogens belong to the genera Borrelia, Rickettsiae, Anaplasma/Ehrlichia (including "Candidatus Neoehrlichia"), Babesia, and Coxiella. The pathogens with the highest detected prevalence were spirochetes of the Borrelia burgdorferi sensu lato complex, in 142 ticks (25.6%). Borrelia afzelii (80/142) was the most frequently detected species, followed by Borrelia burgdorferi sensu stricto (38/142) and Borrelia valaisiana (36/142). Borrelia garinii/Borrelia bavariensis, Borrelia lusitaniae, and Borrelia spielmanii were found in 28 ticks, 5 ticks, and 1 tick, respectively. Rickettsia spp. were detected in 93 ticks (16.8%): R. helvetica (39/93), R. raoultii (38/93), R. monacensis (2/ 93), and R. slovaca (1/93). Thirteen Rickettsia samples remain uncharacterized. "Candidatus Neoehrlichia mikurensis," Babesia spp. (B. venatorum, B. divergens, B. microti), and Anaplasma phagocytophilum were found in 4.5%, 2.7%, and 0.7%, respectively. Coxiella burnetii was not detected. Multiple microorganisms were detected in 40 ticks (7.2%), and the cooccurrence of Babesia spp. and "Candidatus Neoehrlichia mikurensis" showed a significant positive correlation. We also compared different PCRRLBs for detection of Borrelia burgdorferi sensu lato and Rickettsia spp. and showed that different detection approaches provide highly diverse results, indicating that analysis of environmental samples remains challenging. IMPORTANCE This study determined the wide spectrum of tick-borne bacterial and protozoal pathogens that can be encountered in Austria. Surveillance of (putative) pathogenic microorganisms occurring in the environment is of medical importance, especially when those agents can be transmitted by ticks and cause disease. The observation of significant coinfections of certain microorganisms in field-collected ticks is an initial step to an improved understanding of microbial interactions in ticks. In addition, we show that variations in molecular detection methods, such as in primer pairs and target genes, can considerably influence the final results. For instance, detection of certain genospecies of borreliae may be better or worse by one method or the other, a fact of great importance for future screening studies.KEYWORDS Austria, Babesia, Borrelia burgdorferi, Ixodes ricinus, method comparison, reverse line blot, Rickettsia, tick screening T icks carry and transmit a variety of pathogens (bacteria, protozoa, viruses) that can lead to serious infections in humans (1). In Europe, the majority of vector-borne diseases are caused by tick-borne pathogens; thus, knowledge about such pathogens is of medical relevance.The hard tick Ixodes ricinus is the most important vector in Europe (2) and the most

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Section: Discussionmentioning

confidence: 99%

Approaches for Reverse Line Blot-Based Detection of Microbial Pathogens in Ixodes ricinus Ticks Collected in Austria and Impact of the Chosen Method

Schötta

Wijnveld

Stockinger

et al. 2017

Appl Environ Microbiol

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“…Especially in late manifestations of Lyme borreliosis such as ACA the number of organisms in the tissue is low and culture is often not successful. Molecular techniques based on PCR are fast, more sensitive than culture and highly specific . The results of our study have confirmed from formalin‐fixed, paraffin‐embedded lesional tissue that B .…”

Section: Discussionmentioning

confidence: 99%

“…Two independent PCR assays were used to detect B . burgdorferi ‐specific DNA fragments targeting ospA and ospC genes according to Brandt et al All PCR products (ospA, ospC fragments) were analyzed in 2% agarose gels, purified (MiniElute PCR Purification Kit, Qiagen) and subsequently sequenced (BigDyeTerminator V. 3.1 Cycle Sequencing Kit; Applied Biosystems, Darmstadt, Germany; ABI310 genetic analyser, Applied Biosystems). Sequences were further processed using BioEdit version 5.0.9 and identified using National Center for Biotechnology Information‐Basic Local Alignment Search Tool (NCBI‐BLAST).…”

Section: Methodsmentioning

confidence: 96%

“…In a previous study, we could show that assays targeting genes for outer‐membrane lipoproteins ( ospA and ospC ) located on the linear plasmid of B . burgdorferi sl enable identification of Borrelia strains in formalin‐fixed biopsy specimens . Here we aimed to identify Borrelia genotypes in a larger number of biopsies of ACA using ospA and ospC polymerase chain reaction (PCR) assays and to reassess histopathologic and immunohistochemical features of ACA in correlation with genotypes.…”

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confidence: 99%

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Histopathology and immunophenotype of acrodermatitis chronica atrophicans correlated with ospA and ospC genotypes of Borrelia species

et al. 2015

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With assays targeting ospA and ospC genes we confirmed from paraffin-embedded biopsies that B. afzelii, serotype 2, osp C groups Af5, Af2 and Af6 is the main cause of ACA. Specimens commonly showed a combination of band-like T-cell-rich infiltrates with interstitial granulomatous features, a pattern previously under-recognized in ACA. This finding was particularly typical for lesions infected with ospC genotype Af5.

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“…Molecular approaches such as polymerase chain reaction (PCR) can aid in the detection of Borrelia species. A study performed by Brandt et al, showed that Borrelia genotyping is possible from formalin‐fixed paraffin‐embedded tissue. They developed PCR assays targeting the intergenic spacer (IGS), ospA and ospC , followed by sequencing and phylogenetic analysis with a sensitivity up to 85% .…”

Section: Discussionmentioning

confidence: 99%

The many masks of cutaneous Lyme disease

et al. 2015

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Early cutaneous Lyme disease, erythema migrans, may show different histopathologic patterns. The intent of this case series is to raise awareness of these findings to prevent misdiagnosis and keep this entity in the differential. Erythema migrans develops after a tick bite and subsequent infection with the spirochete, Borrelia burgdorferi. It most commonly manifests as a solitary, annular lesion with a bull's-eye appearance. Classic histopathologic findings include superficial and deep perivascular and interstitial lymphocytic infiltrates mixed with plasma cells and eosinophils. We identified and reviewed eight cases of early erythema migrans. Each patient had confirmed B. burgdorferi IgM seropositivity and IgG seronegativity. Histopathologic evaluation of these biopsies reveals a diversity of patterns. Seven of eight cases show sparse to mild perivascular and interstitial mixed infiltrate of variable amount of lymphocytes, eosinophils, neutrophils and plasma cells, with only one case showing a dense inflammatory infiltrate. Epidermal changes such as spongiosis and interface change are seen in some cases. Additionally, perineural lymphocytic infiltrate is seen in one case, periadnexal infiltrate in four cases and pigment incontinence in one case. Based on variable histopathologic findings, it is important to consider erythema migrans in the differential diagnosis for prompt diagnosis and treatment.

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Genotyping ofBorreliafrom formalin‐fixed paraffin‐embedded skin biopsies of cutaneous borreliosis and tick bite reactions by assays targeting the intergenic spacer region,ospAandospCgenes

Abstract: Genotyping of Borrelia can be easily implemented in a routine dermatopathology setting, especially as a fast method to confirm early cutaneous borreliosis. Genotyping could also enable earlier treatment of patients infected with invasive strains.

Cited by 18 publications

References 51 publications

Approaches for Reverse Line Blot-Based Detection of Microbial Pathogens in Ixodes ricinus Ticks Collected in Austria and Impact of the Chosen Method

Approaches for Reverse Line Blot-Based Detection of Microbial Pathogens in Ixodes ricinus Ticks Collected in Austria and Impact of the Chosen Method

Histopathology and immunophenotype of acrodermatitis chronica atrophicans correlated with ospA and ospC genotypes of Borrelia species

The many masks of cutaneous Lyme disease

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