Brucellosis is a zoonosis with non-specific clinical symptoms involving multiple systems and organs. Its prevalence is low in most of EU countries, which can lead to the difficulties in laboratory and clinical diagnostic. Due to its relationship to the Ochrobactrum spp., it may be misclassified in rapid identification systems. We present a case of a 13-year-old immunocompetent girl who was examined several times for fever, fatigue, night sweats and weight loss; laboratory results showed mildly elevated C-reactive protein, anaemia and leukopenia. Four weeks before the onset of symptoms, she had been on a family holiday in Egypt. Given her symptoms, a haemato-oncological or autoimmune disease was considered more likely. The diagnosis of Brucella spondylitis was made after 4 months. The main reasons for this delay were as follows: low specificity of clinical symptoms, delay in completing the travel history, inconclusive initial serological results and misidentification of the blood culture isolate as Ochrobactrum sp. Even in countries with a low incidence of brucellosis, it is essential to educate healthcare professionals about the disease. Low specificity of symptoms and limited experience of laboratory staff may lead to late diagnosis with risk of complications and poor outcome. If Ochrobactrum spp. is detected in clinical specimens by rapid identification, careful re-evaluation must follow and all measures to prevent laboratory-acquired infections must be taken until Brucella spp. is unequivocally excluded.
- MeSH
- bakteriemie * diagnóza mikrobiologie MeSH
- Brucella izolace a purifikace klasifikace MeSH
- brucelóza * diagnóza mikrobiologie MeSH
- chybná diagnóza * MeSH
- gramnegativní bakteriální infekce diagnóza mikrobiologie MeSH
- horečka * mikrobiologie etiologie MeSH
- lidé MeSH
- mladiství MeSH
- Ochrobactrum * genetika izolace a purifikace MeSH
- spondylitida mikrobiologie diagnóza MeSH
- Check Tag
- lidé MeSH
- mladiství MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- kazuistiky MeSH
- Geografické názvy
- Egypt MeSH
Brucellosis is a zoonosis caused by Brucella, which poses a great threat to human health and animal husbandry. Pathogen surveillance is an important measure to prevent brucellosis, but the traditional method is time-consuming and not suitable for field applications. In this study, a recombinase polymerase amplification-SYBR Green I (RPAS) assay was developed for the rapid and visualized detection of Brucella in the field by targeting BCSP31 gene, a conserved marker. The method was highly specific without any cross-reactivity with other common bacteria and its detection limit was 2.14 × 104 CFU/mL or g of Brucella at 40 °C for 20 min. It obviates the need for costly instrumentation and exhibits robustness towards background interference in serum, meat, and milk samples. In summary, the RPAS assay is a rapid, visually intuitive, and user-friendly detection that is highly suitable for use in resource-limited settings. Its simplicity and ease of use enable swift on-site detection of Brucella, thereby facilitating timely implementation of preventive measures.
- MeSH
- Brucella * genetika izolace a purifikace MeSH
- brucelóza * diagnóza mikrobiologie MeSH
- DNA bakterií genetika MeSH
- lidé MeSH
- limita detekce MeSH
- mléko mikrobiologie MeSH
- rekombinasy * metabolismus genetika MeSH
- senzitivita a specificita MeSH
- skot MeSH
- techniky amplifikace nukleových kyselin * metody MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- skot MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Due to limitations in commercial diagnostic methods, this study aimed to develop a reliable real-time polymerase chain reaction (Rt-PCR) assay for early diagnosis of brucellosis. Optimization of the Rt-PCR method was performed on serum samples spiked by Brucella melitensis with different densities ranging from 101 to 108 colony-forming units (cfu)/mL; each density was prepared in ten samples. The limit of detection was investigated by using Thermo DNA extraction kit with Maxima SYBR Green Rt-PCR and two TaqMan probe-based Rt-PCR protocols performed by QuantiTect and TEMPase multiplex PCR master mixes in two thermal cyclers, which were Rotor-Gene and Bio-Rad. The validation of the optimized protocol was carried on 20 brucellosis-negative samples and 20 samples spiked with B. melitensis by using a combination of Thermo DNA extraction kit with TEMPase PCR master mix. SYBR Green Rt-PCR yielded positive results on all samples having ≥ 104 cfu/mL of B. melitensis in both thermal cyclers. Its limit of detection was 112 DNA copies per reaction. The positivity of both probe-based Rt-PCR protocols was 100% and 80% on the samples having 103 cfu/mL and 102 cfu/mL of B. melitensis, respectively. The limit of detection of probe-based protocols was defined as 4 DNA copies per reaction. The optimized Rt-PCR protocol showed high-level accuracy, precision, specificity, and sensitivity, each having a rate of 100%. The current study indicated that the TaqMan probe-based Rt-PCR protocol optimized and validated with serum samples can be reliably used for early diagnosis of brucellosis.
Brucelóza je málo častá, ale závažná bakteriální zoonóza. Česká republika patří mezi státy oficiálně prosté brucelózy u hospodářských zvířat. Na našem území se mohou vyskytnout sporadické importované nákazy člověka způsobené např. Brucella melitensis. V populaci divoce žijících zajíců se však stále vyskytují ojedinělé případy infekce způsobené Brucella suis, která může nakazit i člověka. Stručné sdělení informuje o výskytu dvou případů brucelózy zajíců na Moravě, včetně zkušeností s identifikací izolátů metodou MALDI-TOF.
Brucellosis is a rare but serious bacterial zoonosis. Officially, the Czech Republic is among countries that are free from brucellosis in livestock. In the country, sporadic imported human infections may occur, caused by e.g. Brucella melitensis. In wild hare populations, however, rare cases of infection caused by Brucella suis are still observed, potentially threatening humans. The short communication reports two cases of hare brucellosis in Moravia, Czech Republic, and experiences with isolate identification by the MALDI-TOF method.
- Klíčová slova
- MALDI-TOF,
- MeSH
- Brucella suis MeSH
- brucelóza mikrobiologie přenos veterinární MeSH
- zajíci MeSH
- zoonózy přenos MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- zvířata MeSH
- Geografické názvy
- Česká republika MeSH
Bakteriální zoonózy představují i v současnosti závažný medicínský problém. K méně častým, ale závažným zoonózám patří maltská horečka způsobená bakterií Brucella melitensis. Předkládaná kazuistika popisuje případ relapsu importované maltské horečky u mladého muže. Diagnóza byla vedle čtyř sérologických testů verifikována přímým průkazem původce v hemokultuře. Izolát Brucella melitensis byl identifikován metodou polymerázové řetězové reakce a metodou hmotností spektrometrie MALDI-TOF.
Bacterial zoonoses still represent a serious medical problem. One of the less frequent but severe zoonoses is brucellosis caused by the bacterium Brucella melitensis. The presented case report describes relapsing imported brucellosis in a young male. In addition to four serological tests, the diagnosis was confirmed by direct detection of the pathogen in blood culture. The isolate of Brucella melitensis was identified using the MALDI-TOF BioTyper method and subsequently also by PCR.
- Klíčová slova
- MALDI-TOF, alimentární infekce, zoonóza,
- MeSH
- Brucella melitensis izolace a purifikace MeSH
- brucelóza diagnóza mikrobiologie MeSH
- cestování MeSH
- lidé MeSH
- mladý dospělý MeSH
- Check Tag
- lidé MeSH
- mladý dospělý MeSH
- mužské pohlaví MeSH
- Publikační typ
- kazuistiky MeSH
From the mandibular lymph nodes of wild red foxes (Vulpes vulpes) hunted in the region of Gmünd, Lower Austria, two gram-negative, oxidase- and urease-positive, coccoid rod-shaped bacteria (strains 257 and 284) were isolated. Cells were fast growing, nonmotile, and agglutinated with monospecific anti-Brucella (M) serum. Both strains were biochemically identified as Ochrobactrum anthropi by using the API 20NE test. However, sequencing of the 16S rRNA and recA genes clearly identified strains 257 and 284 as Brucella spp. Further molecular analysis by omp2a/b gene sequencing, multilocus sequence typing and multilocus variable number tandem repeats analysis revealed Brucella microti, a recently described Brucella species that has originally been isolated from diseased common voles (Microtus arvalis) in South Moravia, Czech Republic in 2000. Our findings demonstrate that B. microti is prevalent in a larger geographic area covering the region of South Moravia and parts of Lower Austria. Foxes could have become infected by ingestion of infected common voles.
- MeSH
- Brucella klasifikace genetika izolace a purifikace MeSH
- brucelóza epidemiologie mikrobiologie veterinární MeSH
- genotyp MeSH
- lišky * MeSH
- lymfatické uzliny mikrobiologie MeSH
- mandibula MeSH
- Ochrobactrum anthropi klasifikace izolace a purifikace MeSH
- proteiny vnější bakteriální membrány genetika MeSH
- RecA-rekombinasy genetika MeSH
- RNA ribozomální 16S genetika MeSH
- sekvenční analýza MeSH
- zeměpis MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Česká republika MeSH
- Rakousko MeSH
Autoři předkládají kazuistiku 44 letého pacienta přijatého na infekční oddělení nemocnice Pardubice pro horečnatou lymfadenitidu nejasného původu. Před přijetím pacient léčen na interním oddělení dvojkombinací antibiotik (OXA + GEN). Pro podezření na zoonózu (epidemiologická a pracovní anamnéza, klinický průběh) provedena změna léčby (DOX + GEN), při které dochází promptně k regresi palpačního nálezu na uzlinách a k normalizaci teploty. Opakovaným sérologickým vyšetřením vyloučena ulceroglandulární forma tularémie. Opakovaně zjištěny signifikantní titry protilátek proti Brucella sp. Kazuistika doplněna o údaje dokumentující výskyt brucelózy v CR v letech 1955-1999 u lidí a v letech 1992-2000 u zvířat.
Authors present a case history of a 44-year old patient admitted to Department of Infectious Diseases at Pardubice Hospital with fibrile lymphadenitis of unknown origin. Prior to admission the patient was treated at Internal Medical Department by combination of antibiotics (OXA + GEN). Based on suspicion of zoonosis (epidemiological and work history, clinical course) the treatment was changed (DOX + GEN), resulting in prompt regression of palpation fading on lymf nodes and in normaUzation of body temperature. Repeated serological examination excluded ulceroglandular formo f tularemia. Significant titers of antibodies against Brucella sp. Were proved repeatedly. Case history is completed with data documenting the incidence of brucellosis in the Czech Republic within years 1955-1999 in humus and within 1992-2000 in animals.
- MeSH
- brucelóza mikrobiologie terapie MeSH
- lymfadenitida etiologie terapie MeSH
- zoonózy mikrobiologie MeSH
- Check Tag
- mužské pohlaví MeSH
- Publikační typ
- kazuistiky MeSH
