Restriction fragment length polymorphisms of the DNA of selected Naegleria and Acanthamoeba amebae

McLaughlin, Gerald L.; Brandt, Floy H.; Visvesvara, G. S.

doi:10.1128/jcm.26.9.1655-1658.1988

Cited by 58 publications

(24 citation statements)

References 13 publications

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“…In the absence of any clear-cut morphologic differences between N. fowleri and other Naegleria spp., isoenzyme and DNA restriction fragment length polymorphism (RFLP) analysis or animal pathogenicity tests should be made to differentiate between pathogenic N. fowleri and other Naegleria spp. [17,18]. Both of these procedures require growing large numbers of organisms, which is time-consuming and fastidious.…”

Section: Discussionmentioning

confidence: 99%

Differentiation of Naegleria fowleri and other naegleriae by polymerase chain reaction and hybridization methods

Sparagano¹

1993

FEMS Microbiology Letters

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In order to detect and identify Naegleria fowleri strains an assay based on the Polymerase Chain Reaction (PCR) was evaluated. The amplified DNA fragments were detected by gel electrophoresis and ethidium bromide staining, followed by Southern blot hybridization with an internal digoxigenin-labeled probe. A set of primers (B1B2) which flank a 678-bp region within a virulence-associated gene, allowed for the highly specific identification of N. fowleri, since Naegleriae (N. lovaniensis, N. australiensis, N. gruberi, N. andersoni and N. jadini) and other Protozoa did not react. These primers did not detect amplification products from various organisms: Gram-positive bacteria, Gram-negative bacteria, algae, yeasts and human DNA. Whereas a second set of primers (A1A2), which flank a different sequence, detected various Naegleriae and Acanthamoebae strains. After 40 amplification cycles, the limit of detection was a single cell (cyst or trophozoite). Thus, the PCR appears to be a rapid and powerful tool for identification and detection of N. fowleri.

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Section: Discussionmentioning

confidence: 99%

Differentiation of Naegleria fowleri and other naegleriae by polymerase chain reaction and hybridization methods

Sparagano¹

1993

FEMS Microbiology Letters

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“…Current methods for detection and enumeration of Naegleria species are based on culture techniques 30 followed by various other methods (species-specific monoclonal antibodies 31 , PCR 32 , enzyme electrophoresis 27 , isoenzyme electrophoretic profiles 33,34 and DNA restriction fragment length polymorphisms (RFLPs) 32,35,36 to identify the isolate up to the species level. Randomly amplified polymorphic DNA typing has been used to differentiate Naegleria spp.…”

Section: Discussionmentioning

confidence: 99%

A preliminary study on Naegleria species in water bodies of Kurunegala district, Sri Lanka

Gunarathna

Iddawela

Wickramasinghe³

2018

Sri Lankan J Infec Dis

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Introduction and Objective:Species belonging to the genus Naegleria are free-living ubiquitous protozoa. They have been isolated from most regions of the world. N. fowleri causes an acute, fulminant and rapidly fatal infection involving the central nervous system (CNS) in humans. It is known as primary amoebic meningoencephalitis (PAM). Infection is generally acquired while swimming, diving and total submersion for bathing in freshwater-lakes and ponds. Many inland fresh water bodies are present in Sri Lanka. These water bodies are frequently used by people for their daily needs. However, studies have not yet been conducted to determine the prevalence of Naegleria species occurring in local water bodies. The present study was therefore, carried out to isolate Naegleria species from selected water bodies located in four Divisional Secretariat (DS) divisions in the Kurunegala district, Sri Lanka. Methods:Two different sites (clear and turbid water) of each tank were selected for sampling. Two water samples (surface water and deep water) were collected from each site (4 samples from one tank). Altogether, eighty water samples were collected from 20 tanks. Culture, enflagellation test and staining were done to detect Naegleria species. ArcGIS 10.3 and MINITAB (14) software were used for the data analysis. Results:Flagella transformation was observed in 19 (47.5%) surface water samples and 11 (27.5%) deep water samples. Of 20 tanks, 10 were positive for Naegleria species.Conclusions: Findings of the present study suggest that more specific genotyping studies are needed to confirm the presence of pathogenic N. fowleri in the study area.

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“…Restriction fragment length polymorphism (RFLP) analysis was used to characterize the DNA of presumptive N. fowleri isolates. Amoebae contain repetitive, simple DNA which was extracted and analyzed by using the methods of McLaughlin et al (11). Pellets of amoebae (7 x 106) were lysed and digested by incubation in 5 ml of 1% sodium dodecyl sulfate-5 mM EDTA-0.3 mg of proteinase K per ml-50 mM Tris (pH 8.0) for 2 h at 37°C.…”

Section: Methodsmentioning

confidence: 99%

Thermal ecology of Naegleria fowleri from a power plant cooling reservoir

Huizinga

McLaughlin

1990

Appl Environ Microbiol

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The pathogenic, free-living amoeba Naegleria fowleri is the causative agent of human primary amebic meningoencephalitis. N. fowleri has been isolated from thermally elevated aquatic environments worldwide, but temperature factors associated with occurrence of the amoeba remain undefined. In this study, a newly created cooling reservoir (Clinton Lake, Illinois) was surveyed for Naegleria spp. before and after thermal additions from a nuclear power plant. Water and sediment samples were collected from heated and unheated arms of the reservoir and analyzed for the presence of thermophilic Naegleria spp. and pathogenic N. fowleri. Amoebae were identified by morphology, in vitro cultivation, temperature tolerance, mouse pathogenicity assay, and DNA restriction fragment length analysis. N. fowleri was isolated from the thermally elevated arm but not from the ambient-temperature arm of the reservoir. The probability of isolating thermophilic Naegleria and pathogenic N. fowleri increased significantly with temperature. Repetitive DNA restriction fragment profiles of the N. fowleri Clinton Lake isolates and a known N. fowleri strain of human origin were homogeneous.

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Restriction fragment length polymorphisms of the DNA of selected Naegleria and Acanthamoeba amebae

Cited by 58 publications

References 13 publications

Differentiation of Naegleria fowleri and other naegleriae by polymerase chain reaction and hybridization methods

Differentiation of Naegleria fowleri and other naegleriae by polymerase chain reaction and hybridization methods

A preliminary study on Naegleria species in water bodies of Kurunegala district, Sri Lanka

Thermal ecology of Naegleria fowleri from a power plant cooling reservoir

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