scholarly journals Detection of Campylobacter jejuni and Campylobacter coli in Environmental Waters by PCR Enzyme-Linked Immunosorbent Assay

2002 ◽  
Vol 68 (3) ◽  
pp. 1319-1324 ◽  
Author(s):  
A. D. Sails ◽  
F. J. Bolton ◽  
A. J. Fox ◽  
D. R. A. Wareing ◽  
D. L. A. Greenway
Keyword(s):  
Campylobacter Jejuni ◽  
Immunosorbent Assay ◽  
Water Samples ◽  
Enrichment Culture ◽  
Environmental Water ◽  
Environmental Water Samples ◽  
Enzyme Linked Immunosorbent Assay ◽  
Campylobacter Coli ◽  
Elisa Assay ◽  
Bacterial Cells

ABSTRACT A PCR enzyme-linked immunosorbent assay (ELISA) assay was applied to the detection of Campylobacter jejuni and Campylobacter coli in environmental water samples after enrichment culture. Bacterial cells were concentrated from 69 environmental water samples by using filtration, and the filtrates were cultured in Campylobacter blood-free broth. After enrichment culture, DNA was extracted from the samples by using a rapid-boiling method, and the DNA extracts were used as a template in a PCR ELISA assay. A total of 51 samples were positive by either PCR ELISA or culture; of these, 43 were found to be positive by PCR ELISA and 43 were found to be positive by culture. Overall, including positive and negative results, 59 samples were concordant in both methods. Several samples were positive in the PCR ELISA assay but were culture negative; therefore, this assay may be able to detect sublethally damaged or viable nonculturable forms of campylobacters. The method is rapid and sensitive, and it significantly reduces the time needed for the detection of these important pathogens by 2 to 3 days.

An Enzyme-Linked ImmunoSorbent Assay (ELISA) for the Identification of Naegleria fowleri in Environmental Water Samples

2003 ◽  
Vol 50 (2) ◽  
pp. 109-113 ◽  
Author(s):  
FABIENNE L. REVEILLER ◽  
MARIE-PIERRE VARENNE ◽  
CLAIRE POUGNARD ◽  
PIERRE-ANDRE CABANES ◽  
EMMANUELLE PRINGUEZ ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Water Samples ◽  
Environmental Water ◽  
Environmental Water Samples ◽  
Enzyme Linked Immunosorbent Assay ◽  
Naegleria Fowleri ◽  
Nægleria Fowleri

Production and Characterization of a Monoclonal Antibody to Campylobacter jejuni

2009 ◽  
Vol 72 (4) ◽  
pp. 870-875 ◽  
Author(s):  
S. A. HEO ◽  
R. NANNAPANENI ◽  
M. G. JOHNSON ◽  
J. S. PARK ◽  
K. H. SEO
Keyword(s):  
Monoclonal Antibody ◽  
Campylobacter Jejuni ◽  
Immunosorbent Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Campylobacter Coli ◽  
Carbonate Buffer ◽  
Horse Blood ◽  
Microaerobic Conditions ◽  
Immunoglobulin Subclass

Campylobacter species are a group of spiral-shaped bacteria that can cause disease in humans and animals. We developed a high-affinity monoclonal antibody (MAb) probe that recognizes Campylobacter jejuni cells. Cell suspensions grown under microaerobic conditions at 42°C for 20 h on Bolton agar plates with lysed horse blood were used as live and heat-killed preparations, centrifuged at 8,000 × g for 20 min, and resuspended in carbonate buffer (pH 9.6) for coating on the enzyme-linked immunosorbent assay plates. BALB/c mice were immunized with C. jejuni sonicated cells at 107 CFU/ml to generate MAb-producing hybridoma clones. Of about 500 initial hybridoma clones, MAb 33D2, which reacted with C. jejuni and Campylobacter coli, was selected for further evaluation. MAb 33D2 is in the immunoglobulin subclass G2a and had relatively weaker reactivity with the C. coli strains tested. MAb 33D2 did not show any cross-reactions with the nine non-Campylobacter bacteria tested in the enzyme-linked immunosorbent assay and had a stronger affinity for C. jejuni as live versus heat-killed cells. In Western blot assays, MAb 33D2 recognized two major antigens of 62 and 43 kDa in extracts from C. jejuni cells but only one antigen of 62 kDa in extracts from C. coli cells.

Improving Mycobacterium spp. detection in water: a new methodological proposal

2004 ◽  
Vol 4 (2) ◽  
pp. 103-106
Author(s):  
R. Santos ◽  
S. Gonçalves ◽  
F. Macieira ◽  
F. Oliveira ◽  
R. Rodrigues ◽  
...  
Keyword(s):  
Water Samples ◽  
Recovery Rate ◽  
Opportunistic Infections ◽  
Environmental Water ◽  
Environmental Water Samples ◽  
Sample Treatment ◽  
Detection Time ◽  
Fast Detection ◽  
Enrichment Step ◽  
Mycobacterium Spp

In recent years, non-tuberculous mycobacteria (NTM), once considered merely environmental saprophytes, have emerged as a major cause of opportunistic infections. There is no evidence of human-to-human transmission but they have been found in several environmental water samples. It is, therefore, of the utmost importance to develop methods of rapidly and accurately detecting non-tuberculous mycobacteria in water samples. To obtain a maximum recovery rate and a reduction of Mycobacterium spp. detection time in water samples, different decontamination, enrichment procedures and antibiotics supplements were tested before the inoculation into the Bactec® system. The proposed method of sample treatment (decrease in the decontamination time, followed for a peptone pre-enrichment step and an aztreonam and cefepime supplement) before the inoculation into the Bactec® system proved to be a good option for reliable and fast detection of Mycobacterium spp. in water samples.

The Occurrence of Male-Specific and Somatic Bacteriophages in Polluted South African Waters

1991 ◽  
Vol 24 (2) ◽  
pp. 251-254 ◽  
Author(s):  
R. Kfir ◽  
P. Coubrough ◽  
W. O. K. Grabow
Keyword(s):  
South African ◽  
Water Samples ◽  
Biological Treatment ◽  
Environmental Water ◽  
Environmental Water Samples ◽  
Resistance Pattern ◽  
Male Specific

The occurrence of somatic (F') and male-specific (F') coliphages and Salmonella phages in a variety of environmental water samples was studied using different bacterial hosts. The number of plaque-forming units (pfu) of the different bacteriophages were compared and their resistance pattern to a biological treatment (humus tank) and chlorination was evaluated. The presence of the bacteriophages in shellfish was also studied. The morphology of isolate bacteriophages was examined as well as the visibility of the different plaques formed. Coliphages were found to produce larger and clearer plaques than all other bacteriophages studied. In most of the environmental water samples coliphages outnumbered all other bacteriophages, with the exception of dam water in which higher levels of F' Salmonella phages were detected. The majority of the F' Salmonella phages were shown to be RNA bacteriophages.

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