RESUMO
The increasing threat of Streptococcus gallolyticus subsp. gallolyticus (SGG) infections has gained considerable attention for its strong association with colorectal cancer (CRC). Herein, we proposed real-time fluorescence loop-mediated isothermal amplification (LAMP) as a novel, simple, rapid, and highly sensitive assay for identifying SGG for the first time. This assay was capable of detecting SGG with initial DNA concentrations ranging from 102 to 108 copies per microliter, under isothermal conditions within 30 min via real-time fluorescence monitoring. Our method was tested for specific identification of SGG strains without cross-reaction with other Streptococcus gallolyticus subspecies and Escherichia coli. The developed LAMP shows a superior performance with shorter time and higher sensitivity compared with conventional polymerase chain reaction (PCR). Significantly, this proposed approach was successfully applied for detecting SGG in clinical urine samples, which is non-invasive diagnosis, showing excellent accuracy and reliability to discriminate healthy controls and CRC patients. For comparison, these samples were also tested against PCR assay. These results yielded an analytical sensitivity of 100% and a specificity of 100% for SGG testing using LAMP. The findings suggest LAMP can be employed for detecting SGG infections which is useful for diagnosis and screening of CRC.
Assuntos
Neoplasias Colorretais/microbiologia , DNA Bacteriano/genética , Técnicas de Amplificação de Ácido Nucleico/métodos , Streptococcus gallolyticus subspecies gallolyticus/genética , Adulto , Idoso , Neoplasias Colorretais/urina , DNA Bacteriano/isolamento & purificação , Feminino , Fluorescência , Humanos , Masculino , Pessoa de Meia-Idade , Técnicas de Amplificação de Ácido Nucleico/economia , Streptococcus gallolyticus subspecies gallolyticus/isolamento & purificação , Fatores de TempoRESUMO
Here, we report a case of neonatal calf meningitis due to Streptococcus gallolyticus subsp. gallolyticus (SGG). Clinical, pathological and microbiological findings were evaluated. API Strep, 16S rRNA gene sequencing, rpoB gene sequencing and sodA gene sequencing were used for the complete identification of SGG. This is the first documented report of neonatal calf meningitis due to SGG in veterinary medicine.
Assuntos
Doenças dos Bovinos/microbiologia , Meningite/veterinária , Streptococcus gallolyticus subspecies gallolyticus/isolamento & purificação , Animais , Animais Recém-Nascidos , Técnicas Bacteriológicas/veterinária , Bovinos , Doenças dos Bovinos/patologia , Doenças dos Bovinos/fisiopatologia , Líquido Cefalorraquidiano/citologia , Líquido Cefalorraquidiano/microbiologia , Masculino , Meningite/microbiologia , Meningite/patologia , Meningite/fisiopatologiaRESUMO
Streptococcus gallolyticus subsp. gallolyticus (Sg) has long been reported to display a strong association with colorectal cancer (CRC). It was recently demonstrated to actively promote the development of CRC, underscoring the importance of Sg in both clinical correlation and functional relevance in CRC. Here we investigated several clinical isolates of Sg in their interactions with human colon cancer cells and in mouse models. Some Sg strains were able to stimulate host cell proliferation (proliferation-promoting Sg, PP-Sg) whereas others were not (non-proliferation-promoting Sg, NP-Sg). PP-Sg strains adhered to colon cancer cells much better than NP-Sg strains, suggesting that close contact between Sg and host cells is important. In mice, PP-Sg is significantly better at colonizing the colon tissues of A/J mice compared to NP-Sg, however this difference was not observed in C57BL/6 mice, suggesting that Sg colonization of mouse colon tissues involves specific interactions between bacterial and host factors on the colonic epithelium. Finally, in an azoxymethane-induced mouse model of CRC, PP-Sg promoted tumor development whereas NP-Sg did not. These findings provide clues to the mechanism underlying the Sg-CRC association and have important implications to clinical studies that aim to correlate Sg with clinical and pathological features of CRC.
Assuntos
Neoplasias Colorretais/microbiologia , Neoplasias Colorretais/patologia , Infecções Estreptocócicas/complicações , Streptococcus gallolyticus subspecies gallolyticus/isolamento & purificação , Streptococcus gallolyticus subspecies gallolyticus/fisiologia , Animais , Adesão Celular , Linhagem Celular Tumoral , Proliferação de Células , Modelos Animais de Doenças , Células Epiteliais/microbiologia , Células Epiteliais/fisiologia , Humanos , Camundongos , Streptococcus gallolyticus subspecies gallolyticus/patogenicidadeRESUMO
BACKGROUND: Bacteremia with Clostridium septicum (CS) and Streptococcus gallolyticus subsp. gallolyticus (SGG) have both been associated with colorectal neoplasms (CRN) and colonoscopic examination is advised, however the differences and similarities in colorectal findings are not well known. METHODS: This is a multicenter, comparative study of patients with CS bacteremia [44 of 664 cases (6.6%) of Clostridium spp.] and SGG bacteremia [257 of 596 cases (44.2%) of S. bovis group], carried out in three hospitals from Spain. Clinical findings related to bacteremia and associated CRN were collected. RESULTS: The main sources of infection were abdominal (77.7%) for CS bacteremia and endovascular (75%) for SGG bacteremia. CS bacteremia was more often associated with malignancies, (72.6% vs. 19.4%) and neutropenia (29.5% vs. 3.1%), and was more acute, with shock at presentation (63.6% vs. 3.9%) and higher 30-day mortality (47.7% vs. 9.7%) compared to SGG (P<0.05 for all). Both, patients with CS and SGG bacteremia often had concomitant CRN (43.1% vs. 49.8%) and most of them presented as occult CRN (73.7% vs. 91.4%; P=0.02). CS cases more often had invasive carcinomas (94.7% vs. 19.5%), location of CRN in the right colon (73.7% vs. 23.4%), larger tumor size (median 7 vs. 1.5cm), and a higher overall CRN related mortality rate (68.4% vs. 7.8%) compared to SGG cases (P<0.05 for all). CONCLUSIONS: Both, CS and SGG bacteremia are associated with occult CRN. CS cases more often had advanced carcinomas than SGG cases, suggesting a distinct temporal association with CRN.
Assuntos
Bacteriemia/complicações , Clostridium septicum/isolamento & purificação , Neoplasias Colorretais/diagnóstico , Neoplasias Colorretais/microbiologia , Streptococcus gallolyticus subspecies gallolyticus/isolamento & purificação , Idoso , Idoso de 80 Anos ou mais , Colonoscopia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Neutropenia/complicações , Sangue Oculto , EspanhaRESUMO
Streptococcus gallolyticus subsp. gallolyticus was identified in humans and animals as commensal of the gut and can act as a causative agent of endocarditis and septicemia. A case-control study was performed to identify yet unknown risk factors for the transmission of this facultative pathogen. The prevalence in the gut of 99 healthy volunteers was determined using real-time polymerase chain reaction resulting in 62.5% S. gallolyticus subsp. gallolyticus positive excrements. Subsequent cultivation offered three isolates and epidemiological analysis based on MLST revealed sequence type (ST) 3 and ST 7, previously detected from bovine and endocarditis patients. These results support the hypotheses of the zoonotic potential of this bacterium. Participant questionnaires were evaluated concerning personal characteristics, nutritional habits and animal contact. Specifically, closer contact between participants and animals influenced the colonization of the human gut significantly and was further affected if volunteers used excrement for the fertilization of plants.