An Eternal Microbe:<i>Brucella</i>DNA Load Persists for Years after Clinical Cure

Vrioni, Georgia; Pappas, Georgios; Priavali, Efthalia; Gartzonika, Constantina; Levidiotou, S.

doi:10.1086/588482

Cited by 86 publications

(79 citation statements)

References 24 publications

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“…The inefficiency of conventional diagnostic methods, particularly serology (20), has coerced laboratories and physicians into applying real-time PCR as a more sensitive and specific tool. Real-time PCR reduces the time required to accomplish the detection and also increases the specificity and sensitivity, and it has the ability to quantify the bacterial DNA load (5,21,22). Quantification of the microbial load is valuable for the diagnosis and control of numerous infectious diseases, such as hepatitis and HIV infection.…”

Section: Discussionmentioning

confidence: 99%

“…Early diagnosis of brucellosis results in early initiation of therapy, which plays a key role in the success of the control and eradication programs; nonetheless, accurate diagnosis requires reliable and sensitive diagnostic tools (4). The intracellular localization of the bacteria and the slow evolution of the disease hamper the usefulness of blood cultures (5). Despite being the gold standard, isolation of brucellae from blood culture or other sterile body fluids has a sensitivity of about 70% (4,6).…”

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confidence: 99%

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Efficient Diagnosis and Treatment Follow-Up of Human Brucellosis by a Novel Quantitative TaqMan Real-Time PCR Assay: a Human Clinical Survey

et al. 2014

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cRapid and effective diagnosis of brucellosis is a challenge for clinicians. Even when diagnosis is on time and therapy is initiated, meticulous follow-up appointments are crucial for ensuring the efficacy of the treatment. Due to shortcomings of serological methods, molecular diagnosis, especially real-time PCR, is becoming a main approach in laboratory diagnostics. Thus, the development of efficient procedures and standardization of the PCR tests will have a great impact on the precise detection and quantification of bacterial DNA loads, which is valuable for the medical management of brucellosis patients. We developed a new TaqMan real-time PCR directed to bcsp31, a shared gene of the brucellae. The bcsp31 gene fragment was cloned into pJET1.2. Recombinant pJET1.2-bcsp31 was linearized by HindIII digestion, and the product was used for the preparation of a standard curve. A panel of Brucella spp. and non-Brucella pathogens was tested. No bacterial genomes other than those of the brucellae were detected. According to the results, specificity of the method was 100%. In a clinical assessment, the positive-control group comprised 37 patients with microbiologically confirmed brucellosis, and 25 healthy individuals served as the negative-control group. By the end of the treatment period, there was a significant decrease in the DNA load of the 37 brucellosis patients, which persisted for the 4 weeks of monitoring after treatment, suggesting that our proposed method is an efficient monitoring tool. Serum samples prior to any treatment were collected from the 25 serologically suspicious patients and assessed by our method; 72% of these patients tested positive for brucellosis.T he brucellae are facultative, intracellular, Gram-negative bacteria that infect animals and humans (1). Although it has been eradicated in some developed countries, brucellosis still remains a major problem worldwide. Diagnosis of the disease presents a major challenge to clinical and veterinary services (2, 3). Brucella spp. are able to survive within host cells, leading to relapses and focal complications (1). Early diagnosis of brucellosis results in early initiation of therapy, which plays a key role in the success of the control and eradication programs; nonetheless, accurate diagnosis requires reliable and sensitive diagnostic tools (4). The intracellular localization of the bacteria and the slow evolution of the disease hamper the usefulness of blood cultures (5). Despite being the gold standard, isolation of brucellae from blood culture or other sterile body fluids has a sensitivity of about 70% (4, 6). Serodiagnostic assays are principally based on antibodies against the Brucella lipopolysaccharide (LPS) and have high sensitivity but low specificity. The low specificity may be due to the crossreaction of antibodies with LPS from other bacterial species and subsensitive or high-immunity reactions, depending on the subclinical or endemic prevalence of the disease (4, 6, 7). The presence of IgG and IgM for months after therapy complicates...

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Section: Discussionmentioning

confidence: 99%

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confidence: 99%

Efficient Diagnosis and Treatment Follow-Up of Human Brucellosis by a Novel Quantitative TaqMan Real-Time PCR Assay: a Human Clinical Survey

et al. 2014

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“…A la fecha, la RPC es útil en el diagnóstico de pacientes con brucelosis aguda, como lo observaron Vrioni y cols. Sin embargo, ellos también observaron que de 39 pacientes, 87% seguían teniendo RPC positiva al final de tratamiento, en 77% permanecía positiva después de 6 meses del tratamiento mientras que 70% permaneció positivo al RPC aún después de 2 años; ninguno de los pacientes mostró sintomatología de recaídas 37 . Al inicio de este seguimiento, todos los pacientes tuvieron RPC positiva, mientras que al final sólo aquellos asintomáticos y con serología negativa tuvieron resultados de RPC negativos (el padre, la tía, y la prima).…”

Section: Wwwsochinfclunclassified

“…Además, la presencia de la bacteria en sangre es transitoria, y en otros fluidos la bacteria podría simplemente no estar presente en un nú-mero suficiente que permitiera ser detecta en el momento del análisis. Por otro lado, la presencia de ADN detectable mediante RPC no necesariamente prueba la viabilidad bacteriana ni que la enfermedad esté activa 37 . A pesar de que la RPC es una técnica con un gran valor diagnóstico, la estandarización de la preparación de la muestra, el método de extracción de ADN así como la elección de la secuencia genética a ser amplificada influyen en la interpretación de los resultados.…”

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Seguimiento clínico, serológico y mediante la reacción de polimerasa en cadena de una familia con brucelosis

Morales-García¹,

García-Méndez²,

Regalado-Jacobo³

et al. 2014

Rev. chil. infectol.

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“…In HIV infected patients, as the disease progresses, the number of CD 4 + cells decrease, which leads to cellular immune deficiency (13). In brucellosis, elimination of the bacteria from contaminated macrophages is very hard (14). Iran is one of the endemic countries for brucellosis (15).…”

Section: Introductionmentioning

confidence: 99%

Seroprevalence of Brucellosis in HIV-Infected Patients in Sanandaj, Iran

Rezaee

Rashidi

Ghaedi

et al. 2013

Arch Clin Infect Dis

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Background: Infection with human immunodeficiency virus (HIV) leads to cellular immune deficiency and theoretically patients infected with HIV are susceptible to brucellosis. Objectives: The current study aimed to determine brucellosis rate in the patients infected with HIV. Patients and Methods: We included 89 HIV + patients from Sanandaj Consultation Center for Behavioral Diseases. Patients signed informed written consent before filling out the questionnaire. After serum collection, standard Wright tube, Coombs-Wright and 2ME-Wright tests were performed. Moreover, blood samples obtained from 502 individuals, who were not infected with HIV, were served as the control. Results:The mean age of participants in the experimental and control groups were 33.31 ± 7.47 and 34.38 ± 11.29 years, respectively. In the Wright tube test for the HIV + group, 71 individuals (79.8%) did not have an antibody against Brucella spp., while 18 patients (20.2%) were positive for the antibody. According to the results of Wright tube test for the control group, 63 (12.5%) participants were positive for antiBrucella antibody. The frequency of antibody against Brucella spp. in the HIV + group was significantly higher than that of the control group (P = 0.042). Conclusions: HIV positive individuals in areas endemic for brucellosis must be investigated for the disease.

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An Eternal Microbe:BrucellaDNA Load Persists for Years after Clinical Cure

Abstract: Brucella melitensis DNA persists despite appropriate treatment and apparent recovery. This finding offers a new insight into the pathophysiology of the disease: B. melitensis is a noneradicable, persisting pathogen.

Cited by 86 publications

References 24 publications

Efficient Diagnosis and Treatment Follow-Up of Human Brucellosis by a Novel Quantitative TaqMan Real-Time PCR Assay: a Human Clinical Survey

Efficient Diagnosis and Treatment Follow-Up of Human Brucellosis by a Novel Quantitative TaqMan Real-Time PCR Assay: a Human Clinical Survey

Seguimiento clínico, serológico y mediante la reacción de polimerasa en cadena de una familia con brucelosis

Seroprevalence of Brucellosis in HIV-Infected Patients in Sanandaj, Iran

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