Usefulness of a quantitative real-time PCR assay using serum samples to discriminate between inactive, serologically positive and active human brucellosis

AbstractDiagnosis of brucellosis can be difficult in certain scenarios where conventional microbiological techniques have important limitations. The aim of this study was to develop a LightCycler Quantitative PCR assay in serum samples to discriminate between active and past brucellosis. In total, 110 serum samples from 46 brucellosis patients and 64 controls, including persons who had recently been treated for brucellosis, asymptomatic persons exposed to brucellosis, and patients with febrile syndromes involving a differential diagnosis with brucellosis, were studied. Brucella spp.-specific sequences of the PCR primers and probe were selected from the gene encoding an immunogenic membrane protein of 31 kDa (BCSP31). The analytical sensitivity was 1 × 101 fg of Brucella DNA. The mean threshold cycles for brucellosis patients and controls were 31.8 ± 1.7 and 35.4 ± 1.1, respectively (p <0.001). The best cut-off for bacterial DNA load was 5 × 103 copies/mL. At this cut-off, the area under the receiver operating characteristic curves was 0.963 (95% CI 0.920–1.005), with a sensitivity of 93.5% and a specificity of 98.4%. Under the assay conditions, the LightCycler Quantitative PCR in serum samples seems to be highly reproducible, rapid, sensitive and specific. It is therefore a useful method for both the initial diagnosis and the differentiation between past and active brucellosis

Community-based incidence of acute renal failure

There is limited information about the true incidence of acute renal failure (ARF). Most studies could not quantify disease frequency in the general population as they are hospital-based and confounded by variations in threshold and the rate of hospitalization. Earlier studies relied on diagnostic codes to identify non-dialysis requiring ARF. These underestimated disease incidence since the codes have low sensitivity. Here we quantified the incidence of non-dialysis and dialysis-requiring ARF among members of a large integrated health care delivery system – Kaiser Permanente of Northern California. Non-dialysis requiring ARF was identified using changes in inpatient serum creatinine values. Between 1996 and 2003, the incidence of non-dialysis requiring ARF increased from 322.7 to 522.4 whereas that of dialysis-requiring ARF increased from 19.5 to 29.5 per 100 000 person-years. ARF was more common in men and among the elderly, although those aged 80 years or more were less likely to receive acute dialysis treatment. We conclude that the use of serum creatinine measurements to identify cases of non-dialysis requiring ARF resulted in much higher estimates of disease incidence compared with previous studies. Both dialysis-requiring and non-dialysis requiring ARFs are becoming more common. Our data underscore the public health importance of ARF

Rapid diagnosis of human brucellosis by SYBR Green I-based real-time PCR assay and melting curve analysis in serum samples

SUMMARYThe aim of this study was to develop a LightCycler-based real-time PCR (LC-PCR) assay and to evaluate its diagnostic use for the detection of Brucella DNA in serum samples. Following amplification of a 223-bp gene sequence encoding an immunogenetic membrane protein (BCSP31) specific for the Brucella genus, melting curve and DNA sequencing analysis was performed to verify the specificity of the PCR products. The intra- and inter-assay variation coefficients were 1.3% and 6.4%, respectively, and the detection limit was 5 fg of Brucella DNA (one genome equivalent). After optimisation of the PCR assay conditions, a standard curve was obtained with a linear range (correlation coefficient = 0.99) over seven orders of magnitude from 107 to 10 fg of Brucella DNA. The LC-PCR assay was found to be 91.9% sensitive and 95.4% specific when tested with 65 negative control samples and 62 serum samples from 60 consecutive patients with active brucellosis. The assay is reproducible, easily standardised, minimises the risk of infection in laboratory workers, and has a total processing time of < 2 h. It could therefore form a promising and practical approach for the rapid diagnosis of human brucellosis

A renormalisation group approach to two-body scattering in the presence of long-range forces

We apply renormalisation-group methods to two-body scattering by a combination of known long-range and unknown short-range potentials. We impose a cut-off in the basis of distorted waves of the long-range potential and identify possible fixed points of the short-range potential as this cut-off is lowered to zero. The expansions around these fixed points define the power countings for the corresponding effective field theories. Expansions around nontrivial fixed points are shown to correspond to distorted-wave versions of the effective-range expansion. These methods are applied to scattering in the presence of Coulomb, Yukawa and repulsive inverse-square potentials.Comment: 22 pages (RevTeX), 4 figure

Toward an internally consistent astronomical distance scale

Accurate astronomical distance determination is crucial for all fields in astrophysics, from Galactic to cosmological scales. Despite, or perhaps because of, significant efforts to determine accurate distances, using a wide range of methods, tracers, and techniques, an internally consistent astronomical distance framework has not yet been established. We review current efforts to homogenize the Local Group's distance framework, with particular emphasis on the potential of RR Lyrae stars as distance indicators, and attempt to extend this in an internally consistent manner to cosmological distances. Calibration based on Type Ia supernovae and distance determinations based on gravitational lensing represent particularly promising approaches. We provide a positive outlook to improvements to the status quo expected from future surveys, missions, and facilities. Astronomical distance determination has clearly reached maturity and near-consistency.Comment: Review article, 59 pages (4 figures); Space Science Reviews, in press (chapter 8 of a special collection resulting from the May 2016 ISSI-BJ workshop on Astronomical Distance Determination in the Space Age

ESTUDIO DE CASO DE PROBABLE ENFERMEDAD MUCOSAL EN CIERVOS COLA BLANCA (Odocoileus virginianus)

La enfermedad de las mucosas afecta a bovinos, venados y búfalos, y es ocasionada por el Pestivirus de la familia Falaviviridae, existen 2 biotipos del virus: el citopático (cp) y el no citopático (ncp), según su comportamiento en cultivos celulares. Se presenta el caso de dos ciervos cola blanca (Odocoileus virginianus) remitidos a diagnóstico para estudio anatomopatológico; los animales antes de morir mostraban falta de apetito, postración e incoordinación, lesiones (raspaduras) en la capa y piel de la zona lumbar, ulceraciones en las comisuras de los belfos y nariz. Al estudio anatomopatológico los principales hallazgos observados, fueron un estado de carnes malo, a la inspección interna del tejido subcutáneo en la región submandibular se observó una extensa área de necrosis con exudado fribrinopurulento. En el aparato digestivo fue apreciable la estomatitis con exudado fibrino necrótico y peritonitis; distención ruminal con contenido alimenticio lechoso, abundante material fibroso y grano molido. En aparato urinario se observó congestión renal; y palidez muscular de la región subescapular. Los diagnósticos emitidos fueron: estomatitis fibrino necrótica secundaria de posible etiología mixta, sugestiva de enfermedad mucosal; y miodistrofia de posible etiología nutricional