Investigation of Borrelia burgdorferi genotypes in Australia obtained from erythema migrans tissue

Mayne, Peter

doi:10.2147/ccid.s31913

Cited by 24 publications

(31 citation statements)

References 14 publications

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“…DNA was extracted from the tissue samples using the QIAamp DNA Mini Kit (QIAGEN). The four samples were analyzed in duplicate with positive and negative controls using primers AB-B1 for the Borrelia 16S rRNA gene target, as previously described 21 . The thermal profile for all analyses involved incubation for 2 mins at 50°C, polymerase activation for 10 mins at 95°C then PCR cycling for 40 cycles of 10 secs at 95°C dropping to 60°C sustained for 45 secs.…”

Section: Methodsmentioning

confidence: 99%

Association of spirochetal infection with Morgellons disease

Middelveen¹,

Burugu

Poruri

et al. 2013

F1000Res

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Morgellons disease (MD) is an emerging multisystem illness characterized by skin lesions with unusual filaments embedded in or projecting from epithelial tissue. Filament formation results from abnormal keratin and collagen expression by epithelial-based keratinocytes and fibroblasts. Recent research comparing MD to bovine digital dermatitis, an animal infectious disease with similar skin features, provided clues that spirochetal infection could play an important role in the human disease as it does in the animal illness. Based on histological staining, immunofluorescent staining, electron microscopic imaging and polymerase chain reaction, we report the detection of Borrelia spirochetes in dermatological tissue of four randomly-selected MD patients. The association of MD with spirochetal infection provides evidence that this infection may be a significant factor in the illness and refutes claims that MD lesions are self-inflicted and that people suffering from this disorder are delusional. Molecular characterization of the Borrelia spirochetes found in MD patients is warranted.

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Section: Methodsmentioning

confidence: 99%

Association of spirochetal infection with Morgellons disease

Middelveen¹,

Burugu

Poruri

et al. 2013

F1000Res

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“…Duration > 18 monthsELISA B. burgdorferi s. s. antibody negative.Western blot 2 + antibodies (one level below strongest staining intensity) to outer surface protein A (OspA) of B. garinii only.Biopsy of secondary lesion showed mild, mainly perivascular lymphohistiocytic inflammatory cell infiltrate in superficial dermis, minimal exocytosis, a little pigment incontinence, no organisms demonstrated with PAS, Gram or Warthin–Starry stains.Culture of biopsy grew spirochaetesDirect immunofluorescence antibody staining to flagellin protein of B. burgdorferi s. l. and PCR of the flagellin and 16S rRNA identified B. garinii more closely related to European rather than Asiatic B. garinii strains.Positive/positive (NS)[84]152.8E, 31.66SYesEM, no systemic illness rpoC PCR positiveNP/positive-sequencing showed 99% identity match to B. burgdorferi strain N40[87]152.7E 31.73SNever left AustraliaEM, systemic illness rpoC PCR positiveNP/positive-sequencing showed 99% identity match to B. burgdorferi strain N40[87]151.3E, 33.74SYesEM, fever, meningism, severe headache worse with coughing and shaking of head, photophobia and retro-orbital pain. rpoC PCR positiveFollow-up testing post-treatment revealed: B. burgdorferi IgA, G, M negative. B. burgdorferi multiplex PCR negative (primer targets not specified). Babesia and Bartonella serology negative.NP/positive-sequencing showed 98% identity match to B. burgdorferi strain N40[87]152.8E, 31.32SNever left AustraliaEM, no systemic illnessMultiplex primer set 16S rRNA and OspC used — but only one product sequence is provided. Unclear if both or only one set was positive.NP/positive-sequencing result inconclusive.[87]Rural VictoriaNDFever, regular presumed viral illness, chronic fatigue syndrome. Severe arthritis in hands, auditory hypercusis, poor concentration, irritability and emotional lability, episodic sleep disturbances, two episodes of severe generalized body pain without cause, one episode of auditory hallucinations and paranoid ideas.…”

Section: Lyme-like Case Reported In Australiamentioning

confidence: 99%

Is there a Lyme-like disease in Australia? Summary of the findings to date

2016

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Lyme Borreliosis is a common tick-borne disease of the northern hemisphere caused by the spirochaetes of the Borrelia burgdorferi sensu lato (B. burgdorferi s. l.) complex. It results in multi-organ disease with arthritic, cardiac, neurological and dermatological manifestations. In the last twenty-five years there have been over 500 reports of an Australian Lyme-like syndrome in the scientific literature. However, the diagnoses of Lyme Borreliosis made in these cases have been primarily by clinical presentation and laboratory results of tentative reliability and the true cause of these illnesses remains unknown. A number of animals have been introduced to Australia that may act as B. burgdorferi s. l. reservoirs in Lyme-endemic countries, and there are some Australian Ixodes spp. and Haemaphysalis spp. ticks whose geographical distribution matches that of the Australian Lyme-like cases. Four published studies have searched for Borrelia in Australian ticks, with contradicting results. The cause of the potential Lyme-like disease in Australia remains to be defined. The evidence to date as to whether these illnesses are caused by a Borrelia species, another tick borne pathogen or are due to a novel or unrelated aetiology is summarised in this review.

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“…In samples with sufficient DNA for sequencing, endpoint PCR amplification and Sanger sequencing of the Borrelia gene target from cultures was followed by BLAST comparison with known Borrelia sequences, as previously described ( Mayne et al , 2012). …”

Section: Methodsmentioning

confidence: 99%

Culture and identification of Borrelia spirochetes in human vaginal and seminal secretions

Middelveen¹,

Burke

Sapi

et al. 2015

F1000Res

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Background: Recent reports indicate that more than 300,000 cases of Lyme disease are diagnosed yearly in the USA. Preliminary clinical, epidemiological and immunological studies suggest that infection with the Lyme disease spirochete Borrelia burgdorferi (Bb) could be transferred from person to person via intimate human contact without a tick vector. Failure to detect viable Borrelia spirochetes in vaginal and seminal secretions would argue against this hypothesis. Methods: Patients with and without a history of Lyme disease were selected for the study after informed consent was obtained. Serological testing for Bb was performed on all subjects. Semen or vaginal secretions were inoculated into BSK-H medium and cultured for four weeks. Examination of genital cultures and culture concentrates for the presence of spirochetes was performed using light and darkfield microscopy, and spirochete concentrates were subjected to Dieterle silver staining, anti-Bb immunohistochemical staining, molecular hybridization and PCR analysis for further characterization. Immunohistochemical and molecular testing was performed in three independent laboratories in a blinded fashion. Positive and negative controls were included in all experiments. Results: Control subjects who were asymptomatic and seronegative for Bb had no detectable spirochetes in genital secretions by PCR analysis. In contrast, spirochetes were observed in cultures of genital secretions from 11 of 13 subjects diagnosed with Lyme disease, and motile spirochetes were detected in genital culture concentrates from 12 of 13 Lyme disease patients using light and darkfield microscopy. Morphological features of spirochetes were confirmed by Dieterle silver staining and immunohistochemical staining of culture concentrates. Molecular hybridization and PCR testing confirmed that the spirochetes isolated from semen and vaginal secretions were strains of Borrelia, and all cultures were negative for treponemal spirochetes. PCR sequencing of cultured spirochetes from three couples having unprotected sex indicated that two couples had identical strains of Bb sensu stricto in their semen and vaginal secretions, while the third couple had identical strains of B. hermsii detected in their genital secretions. Conclusions: The culture of viable Borrelia spirochetes in genital secretions suggests that Lyme disease could be transmitted by intimate contact from person to person. Further studies are needed to evaluate this hypothesis.

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Investigation of Borrelia burgdorferi genotypes in Australia obtained from erythema migrans tissue

Cited by 24 publications

References 14 publications

Association of spirochetal infection with Morgellons disease

Association of spirochetal infection with Morgellons disease

Is there a Lyme-like disease in Australia? Summary of the findings to date

Culture and identification of Borrelia spirochetes in human vaginal and seminal secretions

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