Prolonged survival of Bordetella pertussis in a simple buffer after nasopharyngeal secretion aspiration

1992 ◽  
Vol 38 (11) ◽  
pp. 1210-1213 ◽  
Author(s):  
Scott Halperin ◽  
Alicja Kasina ◽  
Margaret Swift
Keyword(s):  
Key Words ◽  
Bordetella Pertussis ◽  
Casein Hydrolysate ◽  
Prolonged Survival ◽  
Nasopharyngeal Aspirate ◽  
Simple Method ◽  
Clinical Specimens ◽  
Laboratory Facilities ◽  
Nasopharyngeal Secretion ◽  
Phosphate Buffered Saline

A simple method for recovery of Bordetella pertussis is described using phosphate-buffered saline containing a casein hydrolysate for transporting secretions collected by nasopharyngeal aspirate. Bordetella pertussis was reisolated from 92% of clinical specimens held at 4 °C for 1 week and from all specimens held at −20 °C. This method will facilitate the centralization of laboratory facilities for the diagnosis of pertussis. Key words: Bordetella pertussis, specimen transport, nasopharyngeal secretions.

scholarly journals Influence of Selective Agents (EMJH-STAFF), Sample Filtration and pH on Leptospira Interrogans Serovar Icterohaemorrhagiae Cultivation and Isolation from Swine Urine

2021 ◽  
Vol 8 (6) ◽  
pp. 90
Author(s):  
Romana Steinparzer ◽  
Tamara Mair ◽  
Christine Unterweger ◽  
Adi Steinrigl ◽  
Friedrich Schmoll
Keyword(s):  
Diagnostic Tests ◽  
Urine Samples ◽  
Epidemiological Surveillance ◽  
Leptospira Interrogans ◽  
Sample Storage ◽  
Clinical Specimens ◽  
Negative Results ◽  
Swine Urine ◽  
Selective Agents ◽  
Phosphate Buffered Saline

Leptospira spp. cause the zoonotic disease leptospirosis, which occurs in numerous mammalians worldwide. Isolation is still important for serotyping and genotyping of Leptospira, which in turn is essential for epidemiological surveillance of leptospirosis and the development of diagnostic tests and vaccines. However, isolation of Leptospira from clinical specimens is inherently insensitive. This study was conducted to examine the influence of selective agents, sample filtration, sample pH and the use of phosphate buffered saline (PBS) buffer for sample storage to improve the success of cultivation and isolation of Leptospira interrogans serovar Icterohaemorrhagiae from swine urine. EMJH (Ellinghausen McCullough, Johnson and Harris) medium including the selective agents sulfamethoxazole, trimethoprim, amphotericin, fosfomycin and 5-fluorouracil (STAFF) increased the success of Leptospira isolation from spiked swine urine samples. Sample filtration yielded only negative results. Isolation in EMJH-STAFF was successful from swine urine with a density as low as 104 Leptospira/mL, and urine with pH ≤ 7 impaired the cultivation rate. Cultivation and isolation were not improved by the addition of PBS to spiked urine samples prior to storage for 24 h at 4 °C. The results of the study demonstrate that cultivation and isolation of leptospires from swine urine can be improved by enhanced methods.

scholarly journals Fast and Sensitive Detection of Bacillus anthracis Spores by Immunoassay

2012 ◽  
Vol 78 (18) ◽  
pp. 6491-6498 ◽  
Author(s):  
Nathalie Morel ◽  
Hervé Volland ◽  
Julie Dano ◽  
Patricia Lamourette ◽  
Patricia Sylvestre ◽  
...  
Keyword(s):  
Bacillus Anthracis ◽  
Electrical Current ◽  
Biological Weapons ◽  
Meso Scale Discovery ◽  
Simple Method ◽  
Content Type ◽  
Bacillus Anthracis Spores ◽  
Luminescent Signal ◽  
Sample Purification ◽  
Phosphate Buffered Saline

ABSTRACTBacillus anthracisis one of the most dangerous potential biological weapons, and it is essential to develop a rapid and simple method to detectB. anthracisspores in environmental samples. The immunoassay is a rapid and easy-to-use method for the detection ofB. anthracisby means of antibodies directed against surface spore antigens. With this objective in view, we have produced a panel of monoclonal antibodies againstB. anthracisand developed colorimetric and electrochemiluminescence (ECL) immunoassays. Using Meso Scale Discovery ECL technology, which is based on electrochemiluminescence (ECL) detection utilizing a sulfo-Tag label that emits light upon electrochemical stimulation (using a dedicated ECL plate reader, an electrical current is placed across the microplate with electrodes integrated into the bottom of the plate, resulting in a series of electrically induced reactions leading to a luminescent signal), a detection limit ranging between 0.3 × 103and 103CFU/ml (i.e., 30 to 100 spores per test), depending on theB. anthracisstrain assayed, was achieved. In complex matrices (5 mg/ml of soil or simulated powder), the detection level (without any sample purification or concentration) was never altered more than 3-fold compared with the results obtained in phosphate-buffered saline.

scholarly journals A simple method for inhibition free PCR amplification of target DNA directly from clinical specimens

1997 ◽  
Vol 12 (1) ◽  
pp. 78-80 ◽  
Author(s):  
Senthilkumar Kamatchiammal ◽  
Dhashinamoorthy Saravanakumar ◽  
Nagalingeswaran Kumarasamy ◽  
Sunithi Solomon ◽  
Manjula Sritharan ◽  
...  
Keyword(s):  
Pcr Amplification ◽  
Simple Method ◽  
Clinical Specimens ◽  
Target Dna

scholarly journals Depletion of Human DNA in Spiked Clinical Specimens for Improvement of Sensitivity of Pathogen Detection by Next-Generation Sequencing

2016 ◽  
Vol 54 (4) ◽  
pp. 919-927 ◽  
Author(s):  
Mohammad R. Hasan ◽  
Arun Rawat ◽  
Patrick Tang ◽  
Puthen V. Jithesh ◽  
Eva Thomas ◽  
...  
Keyword(s):  
Next Generation Sequencing ◽  
Pathogen Detection ◽  
Clinical Samples ◽  
Metagenomic Sequencing ◽  
Next Generation ◽  
Simple Method ◽  
Clinical Specimens ◽  
Human Dna ◽  
Pathogen Dna ◽  
Generation Sequencing

Next-generation sequencing (NGS) technology has shown promise for the detection of human pathogens from clinical samples. However, one of the major obstacles to the use of NGS in diagnostic microbiology is the low ratio of pathogen DNA to human DNA in most clinical specimens. In this study, we aimed to develop a specimen-processing protocol to remove human DNA and enrich specimens for bacterial and viral DNA for shotgun metagenomic sequencing. Cerebrospinal fluid (CSF) and nasopharyngeal aspirate (NPA) specimens, spiked with control bacterial and viral pathogens, were processed using either a commercially available kit (MolYsis) or various detergents followed by DNase prior to the extraction of DNA. Relative quantities of human DNA and pathogen DNA were determined by real-time PCR. The MolYsis kit did not improve the pathogen-to-human DNA ratio, but significant reductions (>95%;P< 0.001) in human DNA with minimal effect on pathogen DNA were achieved in samples that were treated with 0.025% saponin, a nonionic surfactant. Specimen preprocessing significantly decreased NGS reads mapped to the human genome (P< 0.05) and improved the sensitivity of pathogen detection (P< 0.01), with a 20- to 650-fold increase in the ratio of microbial reads to human reads. Preprocessing also permitted the detection of pathogens that were undetectable in the unprocessed samples. Our results demonstrate a simple method for the reduction of background human DNA for metagenomic detection for a broad range of pathogens in clinical samples.

scholarly journals Molecular epidemiology of Bordetella pertussis in Cambodia determined by direct genotyping of clinical specimens

2017 ◽  
Vol 62 ◽  
pp. 56-58 ◽  
Author(s):  
Takumi Moriuchi ◽  
Ork Vichit ◽  
Yong Vutthikol ◽  
Md. Shafiqul Hossain ◽  
Chham Samnang ◽  
...  
Keyword(s):  
Molecular Epidemiology ◽  
Bordetella Pertussis ◽  
Clinical Specimens

Effect of butt joints on the flexural stiffness of laminated timber bridges

1990 ◽  
Vol 17 (5) ◽  
pp. 859-864 ◽  
Author(s):  
Leslie G. Jaeger ◽  
Baidar Bakht
Keyword(s):  
Key Words ◽  
Flexural Rigidity ◽  
Bridge Decks ◽  
Butt Joint ◽  
Flexural Stiffness ◽  
Butt Joints ◽  
Simple Method ◽  
Timber Bridges

Prestressed and nailed laminated timber bridge decks are made from laminates which, because of their being usually shorter in length than the deck span, are butt-jointed at regular intervals. In calculating deflections of such decks, it is usual to ignore the reduction in flexural rigidity of the deck caused by the presence of the butt joint. The effect of butt joints on the flexural rigidity of the laminated deck is studied analytically, and it is shown that the deflections of a deck having such joints may be significantly larger than those of a deck without them. A simple method is presented to account conservatively for the presence of butt joints in the calculation of the deflections of a prestressed laminated timber deck. Key words: timber bridge, laminated deck, butt joint, prestressed wood deck.

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