Microscopic detection of bacillus Calmette-Guérin mycobacteria in bladder biopsy using fluorescence in situ hybridization: Détection microscopique des Bacilles biliés de Calmette et Guérin (BCG) dans une biopsie vésicale par hybridation in situ en fluorescence.

Intravesical instillation of Bacilli Calmette Guérin (BCG) as a superficial bladder cancer treatment is generally well tolerated, but local or systemic complications may occur, some of which may be life-threatening. Following the suspicion of post-BCG cystitis in a 72-year-old man with a history of urothelial carcinoma treated by intravesical BCG instillation, we used fluorescence in situ hybridization (FISH) targeting the rpoB gene of the Mycobacterium tuberculosis complex to detect Mycobacterium bovis BCG in paraffin-embedded bladder biopsy sections. FISH yielded specific detection of BCG mycobacteria in the bladder biopsy section, appearing as red-fluorescent bacilli. Treatment with rifampicin, ethambutol and isoniazid is then initiated in combination with corticosteroid therapy.

18F-FDG brain PET hypometabolism in patients with long COVID.

PURPOSE: In the context of the worldwide outbreak of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), some patients report functional complaints after apparent recovery from COVID-19. This clinical presentation has been referred as "long COVID." We here present a retrospective analysis of 18F-FDG brain PET of long COVID patients from the same center with a biologically confirmed diagnosis of SARS-CoV-2 infection and persistent functional complaints at least 3 weeks after the initial infection. METHODS: PET scans of 35 patients with long COVID were compared using whole-brain voxel-based analysis to a local database of 44 healthy subjects controlled for age and sex to characterize cerebral hypometabolism. The individual relevance of this metabolic profile was evaluated to classify patients and healthy subjects. Finally, the PET abnormalities were exploratory compared with the patients' characteristics and functional complaints. RESULTS: In comparison to healthy subjects, patients with long COVID exhibited bilateral hypometabolism in the bilateral rectal/orbital gyrus, including the olfactory gyrus; the right temporal lobe, including the amygdala and the hippocampus, extending to the right thalamus; the bilateral pons/medulla brainstem; the bilateral cerebellum (p-voxel < 0.001 uncorrected, p-cluster < 0.05 FWE-corrected). These metabolic clusters were highly discriminant to distinguish patients and healthy subjects (100% correct classification). These clusters of hypometabolism were significantly associated with more numerous functional complaints (brainstem and cerebellar clusters), and all associated with the occurrence of certain symptoms (hyposmia/anosmia, memory/cognitive impairment, pain and insomnia) (p < 0.05). In a more preliminary analysis, the metabolism of the frontal cluster which included the olfactory gyrus was worse in the 7 patients treated by ACE drugs for high blood pressure (p = 0.032), and better in the 3 patients that had used nasal decongestant spray at the infectious stage (p < 0.001). CONCLUSION: This study demonstrates a profile of brain PET hypometabolism in long COVID patients with biologically confirmed SARS-CoV-2 and persistent functional complaints more than 3 weeks after the initial infection symptoms, involving the olfactory gyrus and connected limbic/paralimbic regions, extended to the brainstem and the cerebellum. These hypometabolisms are associated with patients' symptoms, with a biomarker value to identify and potentially follow these patients. The hypometabolism of the frontal cluster, which included the olfactory gyrus, seems to be linked to ACE drugs in patients with high blood pressure, with also a better metabolism of this olfactory region in patients using nasal decongestant spray, suggesting a possible role of ACE receptors as an olfactory gateway for this neurotropism.

Unconventional diagnostic tests for Lyme borreliosis: a systematic review.

BACKGROUND: Lyme borreliosis (LB) diagnosis currently relies mainly on serological tests and sometimes PCR or culture. However, other biological assays are being developed to try to improve Borrelia-infection diagnosis and/or monitoring. OBJECTIVES: To analyse available data on these unconventional LB diagnostic assays through a systematic literature review. METHODS: We searched PubMed and Cochrane Library databases according to the PRISMA-DTA method and the Cochrane Handbook for Systematic Reviews of Interventions. We analysed controlled and uncontrolled studies (published 1983-2018) on biological tests for adults to diagnose LB according to the European Study Group for Lyme Borreliosis or the Infectious Diseases Society of America definitions, or identify strongly suspected LB. Two independent readers evaluated study eligibility and extracted data from relevant study reports; a third reader analysed full texts of papers to resolve disagreements. The quality of each included study was assessed with the QUADAS-2 evaluation scale. RESULTS: Forty studies were included: two meta-analyses, 25 prospective controlled studies, five prospective uncontrolled studies, six retrospective controlled studies and two case reports. These biological tests assessed can be classified as: (i) proven to be effective at diagnosing LB and already in use (CXCL-13 for neuroborreliosis), but not enough to be standardized; (ii) not yet used routinely, requiring further clinical evaluation (CCL-19, OspA and interferon-α); (iii) uncertain LB diagnostic efficacy because of controversial results and/or poor methodological quality of studies evaluating them (lymphocyte transformation test, interferon-γ, ELISPOT); (iv) unacceptably low sensitivity and/or specificity (CD57+ natural killer cells and rapid diagnostic tests); and (v) possible only for research purposes (microscopy and xenodiagnoses). DISCUSSION: QUADAS-2 quality assessment demonstrated high risk of bias in 25/40 studies and uncertainty regarding applicability for 32/40, showing that in addition to PCR and serology, several other LB diagnostic assays have been developed but their sensitivities and specificities are heterogeneous and/or under-evaluated or unassessed. More studies are warranted to evaluate their performance parameters. The development of active infection biomarkers would greatly advance LB diagnosis and monitoring.

Lyme borreliosis and other tick-borne diseases. Guidelines from the French scientific societies (II). Biological diagnosis, treatment, persistent symptoms after documented or suspected Lyme borreliosis.

The serodiagnosis of Lyme borreliosis is based on a two-tier strategy: a screening test using an immunoenzymatic technique (ELISA), followed if positive by a confirmatory test with a western blot technique for its better specificity. Lyme serology has poor sensitivity (30-40%) for erythema migrans and should not be performed. The seroconversion occurs after approximately 6 weeks, with IgG detection (sensitivity and specificity both>90%). Serological follow-up is not recommended as therapeutic success is defined by clinical criteria only. For neuroborreliosis, it is recommended to simultaneously perform ELISA tests in samples of blood and cerebrospinal fluid to test for intrathecal synthesis of Lyme antibodies. Given the continuum between early localized and disseminated borreliosis, and the efficacy of doxycycline for the treatment of neuroborreliosis, doxycycline is preferred as the first-line regimen of erythema migrans (duration, 14 days; alternative: amoxicillin) and neuroborreliosis (duration, 14 days if early, 21 days if late; alternative: ceftriaxone). Treatment of articular manifestations of Lyme borreliosis is based on doxycycline, ceftriaxone, or amoxicillin for 28 days. Patients with persistent symptoms after appropriate treatment of Lyme borreliosis should not be prescribed repeated or prolonged antibacterial treatment. Some patients present with persistent and pleomorphic symptoms after documented or suspected Lyme borreliosis. Another condition is eventually diagnosed in 80% of them.

Lyme borreliosis and other tick-borne diseases. Guidelines from the French Scientific Societies (I): prevention, epidemiology, diagnosis.

Lyme borreliosis is transmitted en France by the tick Ixodes ricinus, endemic in metropolitan France. In the absence of vaccine licensed for use in humans, primary prevention mostly relies on mechanical protection (clothes covering most parts of the body) that may be completed by chemical protection (repulsives). Secondary prevention relies on early detection of ticks after exposure, and mechanical extraction. There is currently no situation in France when prophylactic antibiotics would be recommended. The incidence of Lyme borreliosis in France, estimated through a network of general practitioners (réseau Sentinelles), and nationwide coding system for hospital stays, has not significantly changed between 2009 and 2017, with a mean incidence estimated at 53 cases/100,000 inhabitants/year, leading to 1.3 hospital admission/100,000 inhabitants/year. Other tick-borne diseases are much more seldom in France: tick-borne encephalitis (around 20 cases/year), spotted-fever rickettsiosis (primarily mediterranean spotted fever, around 10 cases/year), tularemia (50-100 cases/year, of which 20% are transmitted by ticks), human granulocytic anaplasmosis (<10 cases/year), and babesiosis (<5 cases/year). The main circumstances of diagnosis for Lyme borreliosis are cutaneous manifestations (primarily erythema migrans, much more rarely borrelial lymphocytoma and atrophic chronic acrodermatitis), neurological (<15% of cases, mostly meningoradiculitis and cranial nerve palsy, especially facial nerve) and rheumatologic (mostly knee monoarthritis, with recurrences). Cardiac and ophtalmologic manifestations are very rarely encountered.

Management of patients presenting with generalized musculoskeletal pain and a suspicion of Lyme disease.

Lyme disease is caused by bacteria of the B. burgdorferi sensu lato complex, and can give polymorphic clinical manifestations that can affect several organs such as the skin, the central nervous system, or the joints. In recent years, patients' associations and physicians have been supporting the hypothesis that this infection would manifest as chronic generalized musculoskeletal pain symptoms, named "chronic Lyme disease". Fibromyalgia is a clinical presentation characterized by chronic generalized musculoskeletal pain with a major impact on quality of life and social and psychological functioning. We analyzed existing literature data on pain syndromes associated with Lyme disease (post-treatment Lyme disease syndrome) or tick bites (polymorphic symptoms after a tick bite). We also analyzed existing data on the diagnosis, pathophysiology, and treatment of fibromyalgia. Our review shows that post-treatment Lyme disease syndrome has characteristics very close to post-infectious fibromyalgia. On the other hand, patients presenting for Lyme disease screening because of chronic generalized musculoskeletal pain symptoms after a tick bite should also be screened for fibromyalgia to allow appropriate management. Antibiotics are not recommended here.

Functional neuroimaging in patients presenting with somatoform disorders: A model for investigating persisting symptoms after tick bites and post-treatment Lyme disease syndrome?

Approximately 10% of patients presenting with Lyme disease experience fatigue, musculoskeletal pain, concentration disorders, or short-term memory deficits in the six months following treatment. This entity has been defined as post-Lyme disease syndrome or post-treatment Lyme disease syndrome. The pathophysiology of this syndrome is unknown, but neither persistence of the bacterium nor effectiveness of antibiotics are currently reported in the literature. The French High Council for Public Health (French acronym HCSP) has recently defined a new entity called "persistent polymorphic symptoms after a tick bite" allowing for designing studies to better understand these subjective presentations, for which objective biomarkers are currently lacking. This entity encompasses patients experiencing fatigue and generalized pain in the months following a tick bite and can be associated with several subjective symptoms with major impact on the quality of life. In the field of somatoform disorders, this article reviews functional neuroimaging studies in patients presenting with subjective complaints and discusses potential clinical implications for persisting symptoms after tick bites and post-treatment Lyme disease syndrome.

Limitations of diagnostic tests for bacterial infections.

Lyme disease diagnosis is currently based on serology - an indirect diagnostic method - as laboratory cultures are fastidious. The only direct diagnostic method that can be useful with some specimens (cutaneous biopsies or aspiration fluid) is PCR. We aimed to detail the main limitations of serology and PCR testing in the diagnosis of bacterial infections. Limitations are supported by examples from the recent history of microbiology. The main limitation of bacterial serology is the presence of numerous cross-reactions due to many genes that are common to various bacterial species. Some serological techniques, such as those used for the diagnosis of rickettsioses mainly, have even been based on the existence of cross-reactions. The main limitation of PCR testing is the potential presence of laboratory contaminations. PCR-performing laboratories must therefore be certified for the use of this technique. PCR testing also does not inform on the viability of the identified bacterium and should therefore be interpreted in light of the clinical presentation. These limitations highlight that all diagnostic test results should not be interpreted on their own; the clinical and epidemiological contexts should always be taken into consideration.

Review of European and American guidelines for the diagnosis of Lyme borreliosis.

Lyme disease is a tick-borne bacterial disease with polymorphic clinical manifestations (cutaneous, rheumatological, and neurological). In recent years the issue of the diagnosis of this infection has been highly publicized on the Internet and other media in Europe and America. Some patients and physicians may share the perception that the diagnosis of the infection is not reliable in France. We reviewed current European and American guidelines on Lyme disease and performed a methodological evaluation of all guidelines. We retrieved 16 guidelines from seven countries. Our analysis revealed a global consensus regarding diagnosis at each stage of the infection. All guidelines indicate that the diagnosis is currently based on a two-tier serology at all stages of the infection, except for the early localized dermatological presentation known as Erythema migrans. One text of so-called guidelines has discordant recommendations when compared with the other guidelines, possibly explained by its low quality score. Contrary to the intense debate taking place on the Internet and in the European and American media, our analysis shows that the great majority of medical scientific guidelines with a high quality score, agree on the clinical diagnostic methods of Lyme disease.

Genotyping of Coxiella burnetii detected in placental tissues from aborted dairy cattle in the north of Algeria.

Coxiella burnetii, is an obligate intracellular bacterium which is present throughout the world. In humans, C. burnetii is the causative agent of Q fever. In cattle, the infection is suspected to cause stillbirths, retained fetal membranes, metritis and infertility. The birth products of ruminants shed huge amounts of bacteria, and are considered a major source for human infection. The present study was designed to search for the presence of C. burnetii in placental tissues collected from aborted and normal calving dairy cows in Algeria, using molecular tools. A total of 77 placental tissue fragments were collected from dairy cows. 73 samples were collected from aborted cows and four samples were collected from natural calving cows over a period of two years from January 2013 to March 2015. The presence of C. burnetii in these samples was screened by quantitative real-time polymerase chain reaction (qPCR) targeting two different genes, IS1111 and IS30 A. The positive PCR amplicons were subsequently sequenced for Multispacer Sequence Typing determination (MST) using seven pairs of sequences (Cox2, Cox5, Cox18, Cox37, Cox56, Cox57, and Cox61). Fourteen placental tissues (19.1%) were found to be positive for C. burnetii by qPCR; 9 (12.3%) from the city of Blida and 5 (6.84%) from the city of Medea. Genotyping of the corresponding amplicons displayed 100% identity with C. burnetii MST20 genotype, confirming the circulation of this clone in dairy farms from Algeria.