Enzyme-linked synthetic oligonucleotide probes: non-radioactive detection of enterotoxigenic<i>Escherichia coli</i>in faecal specimens

Li, Peng; Medon, Peter P.; Skingle, Derek C.; Lanser, J.B.K.; Symons, Robert H.

doi:10.1093/nar/15.13.5275

Cited by 87 publications

(26 citation statements)

References 31 publications

(35 reference statements)

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“…Their comparative advantages and disadvantages have been reviewed extensively (Al-Hakim and Hull, 1986;Zwadyk, Cooksey and Thomsberry, 1986;Donovan etal., 1987;Mifflin et al, 1987;Tabares, 1987). Radioactive labels include 3H, 3'S, 32P and 1251, while non-radioactive probes include various biotin-labelling systems (Al-Hakim and Hull, 1986;Dahlen, 1987;Gebeyehu et al, 1987;McInnes et al, 1987), fluorescein label (Zuckermann, Corey and Schultz, 1987), enzyme labels (Li et al, 1987;McLaughlin et al, 1987;Seriwatana etal., 1987) or, more recently, labelling with steroid antigens (Boehringer Mannheim, 1988b;Schafer, Zischler and Epplen, 1988). Radioactive dNTPs are the traditional labels used for hybridization.…”

Section: Nucleic Acid Hybridization Methodsmentioning

confidence: 99%

New Approaches to the Detection of Microbial Plant Pathogens

Chu

Waterhouse

Martín

et al. 1989

Biotechnology and Genetic Engineering Reviews

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Section: Nucleic Acid Hybridization Methodsmentioning

confidence: 99%

New Approaches to the Detection of Microbial Plant Pathogens

Chu

Waterhouse

Martín

et al. 1989

Biotechnology and Genetic Engineering Reviews

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“…Nonspecific binding of biotin-labelled probes resulting in high background is a common problem (Chan et al, 1985;Peng Li et al, 1987), and researchers have used a number of different methodologies in an attempt to lower background. Small sample sizes undoubtedly limited statistical power.…”

Section: Discussionmentioning

confidence: 99%

Assessment of pollution impact on aquatic microbial communities based on DNA hybridization and protozoan identification: preliminary method development and comparison

Palmer

Niederlehner

Cairns

1994

J Aquat Ecosyst Stress Recov

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Common methods of assessing the biological impact of point source discharge involve a census of organisms above and below the discharge. The identification and enumeration of organisms necessary for this approach can be time consuming and costly. When microbial communities are used, identification and enumeration are often impossible because the majority of species are identifiable only when cultured and most native species cannot be cultured. As such, little is known about the response of nonculturable aquatic microbial communities to anthropogenic changes. We isolated aquatic microbial DNA from colonized artificial substrates in two streams receiving industrial waste. DNA was isolated from the microbial communities upstream and downstream of the discharges and the similarity of these communities were compared by DNA-DNA hybridization. These novel estimates of impact were compared to more conventional estimates based on a component of the microbial communities that could be readily identified, the protozoans. Protozoan species were identified and similarity between upstream and downstream stations was assessed using Jaccard's similarity index.In the Roanoke River, protozoan species richness was significantly lower at the downstream station. Both Jaccard's coefficients and DNA hybridizations indicated that upstream communities were more similar to each other than with the downstream site. However, these differences were not significant. Est'mates of community similarity from Jaccard's coefficients and DNA hybridizations were not correlated. In Peak Creek, protozoan species richness was significantly lower at the downstream station. Jaccard's coefficients and DNA hybridizations were not significantly higher within upstream stations than between upstream and downstream stations. The two similarity measures were not correlated. Problems in this preliminary study included small sample sizes and highly variable background on hybridization membranes resulting from the use of nonradioactive DNA probe label and detection methods. Alternative approaches using radioisotopes or COT curves may be more viable means of making molecular methods a useful tool in impact assessment.

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“…There fore, linkers are incorporated during oligonucleotide synthesis and the nonra dioactive labels are attached after synthesis (117). Labels that can be con veniently attached include biotin (117), fluorochromes (40, 117), chemilu minescent moieties (117), and enzymes (alkaline phosphatase, horseradish peroxidase) (51, 67,106,117). In some studies, enzyme-linked oligonuc leotides obtained the same sensitivity as p32-labelled oligonucleotides (67,117), and they may be the most efficient nonradioactive labelling method for oligonucleotides.…”

Section: Nonradioactive Methodsmentioning

confidence: 99%

“…Oligonucleotide probes have been made to specific genes characteristic of the target organism, such as virulence factors in E. coli (67,77,106), Listeria (24), and Yersinia (73), and to a degradative gene in Pseudomonas cepacia (114). A commercially marketed oligonucleotide probe has been made to detect clinically important Mycobacterium avium (26, 59).…”

Section: Oligonucleotide Probesmentioning

confidence: 99%