Differences in a ribosomal DNA sequence of morphologically indistinguishable species within the Hypodontus macropi complex (Nematoda: Strongyloidea)

Chilton, Neil B.; Gasser, Robin B.; Beveridge, Ian

doi:10.1016/0020-7519(94)00171-j

Cited by 271 publications

(136 citation statements)

References 13 publications

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“…Inter-and intra-species differences were evaluated using BioEdit (www.mbio.ncsu.edu/ bioedit) or Geneious software. Levels of sequence diversity (D) in each target were calculated pairwise using the formula D512(M/L) (Chilton et al, 1995), in which M is the number of shared positions and L is the total number of alignment positions compared. Restriction sites and fragment sizes were predicted in silico and appropriate restriction enzymes for RFLP analysis were selected using DNASIS software (Hitachi DNASIS MAX version 3.0).…”

Section: Differentiation Of M Canis and Its Close Relativesmentioning

confidence: 99%

Multilocus differentiation of the related dermatophytes Microsporum canis, Microsporum ferrugineum and Microsporum audouinii

Rezaei‐Matehkolaei

Makimura

Hoog

et al. 2012

Journal of Medical Microbiology

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Microsporum ferrugineum, an uncommon causative agent of dermatophytosis, has restricted endemicity. Iranian strains suspected to be M. ferrugineum from two patients with tinea were analysed using the rDNA internal transcribed spacer (ITS) region and the partial b-tubulin (BT2) and translation elongation factor 1-a (TEF1) genes. Strains were compared to reference strains to differentiate M. ferrugineum from its relatives Microsporum canis and Microsporum audouinii. Inter-species differences for TEF1 and BT2 were found to be higher than for the ITS region, which is the current molecular standard for species identification in dermatophytes. Intra-species variation was zero for each of the markers. In silico analysis showed that the restriction enzymes BanI and BshNI were together sufficient to differentiate the three species based on TEF1, whereas a two-step digestion was needed with BT2 or the ITS region. The prevalence of M. ferrugineum in clinical samples in Iran appeared to be higher than suspected on the basis of routine phenotypic identification.

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Section: Differentiation Of M Canis and Its Close Relativesmentioning

confidence: 99%

Multilocus differentiation of the related dermatophytes Microsporum canis, Microsporum ferrugineum and Microsporum audouinii

Rezaei‐Matehkolaei

Makimura

Hoog

et al. 2012

Journal of Medical Microbiology

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show abstract

“…The pairwise comparisons were made of the level of sequence differences (Chilton et al, 1995). Arlequin v.3.1 (Excoffier et al, 2005) was used to calculate the population pairwise Fst (Distance method), Fst P values, and the matrix of significant Fst P values Table 1 Countries, geographical locations and host species used in this study, together with the number of host animals and Sarcoptes mite samples, followed by GenBank TM accession numbers for ITS-2+ sequences.…”

Section: Molecular Analysismentioning

confidence: 99%

Is ITS-2 rDNA suitable marker for genetic characterization of Sarcoptes mites from different wild animals in different geographic areas?

Alasaad

Soglia

Spalenza

et al. 2009

Veterinary Parasitology

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“…Accordingly, ITS-1 may overcome the problems of DNA and rRNA persistence in inactivated cells. While the ITS region has been used previously to classify and detect organisms (6,16,28,42), it has not been used to determine viability. The goal of this study was to develop a qPCR method to determine the levels of total and viable Ascaris eggs in laboratory solutions using ITS-1 rDNA and rRNA.…”

mentioning

confidence: 99%

A Real-Time PCR Method for Quantifying Viable Ascaris Eggs Using the First Internally Transcribed Spacer Region of Ribosomal DNA

Pecson

Barrios

Johnson

et al. 2006

Appl Environ Microbiol

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Worldwide, 1.4 billion people are infected with the intestinal worm Ascaris lumbricoides. As a result, Ascaris eggs are commonly found in wastewater and sludges. The current microscopy method for detecting viable Ascaris eggs is time-and labor-intensive. The goal of this study was to develop a real-time quantitative PCR (qPCR) method to determine the levels of total and viable Ascaris eggs in laboratory solutions using the first internally transcribed spacer (ITS-1) region of ribosomal DNA (rDNA) and rRNA. ITS-1 rDNA levels were proportional to Ascaris egg cell numbers, increasing as eggs developed from single cells to mature larvae and ultimately reaching a constant level per egg. Treatments causing >99% inactivation (high heat, moderate heat, ammonia, and UV) eliminated this increase in ITS-1 rDNA levels and caused decreases that were dependent on the treatment type. By taking advantage of this difference in ITS-1 rDNA level between viable, larvated eggs and inactivated, single-celled eggs, qPCR results were used to develop inactivation profiles for the different treatments. No statistical difference from the standard microscopy method was found in 75% of the samples (12 of 16). ITS-1 rRNA was detected only in samples containing viable eggs, but the levels were more variable than rDNA levels and ITS-1 rRNA could not be used for quantification. The detection limit of the rDNA-based method was approximately one larvated egg or 90 single-celled eggs; the detection limit for the rRNA-based method was several orders of magnitude higher. The rDNA qPCR method is promising for both research and regulatory applications.

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Differences in a ribosomal DNA sequence of morphologically indistinguishable species within the Hypodontus macropi complex (Nematoda: Strongyloidea)

Cited by 271 publications

References 13 publications

Multilocus differentiation of the related dermatophytes Microsporum canis, Microsporum ferrugineum and Microsporum audouinii

Multilocus differentiation of the related dermatophytes Microsporum canis, Microsporum ferrugineum and Microsporum audouinii

Is ITS-2 rDNA suitable marker for genetic characterization of Sarcoptes mites from different wild animals in different geographic areas?

A Real-Time PCR Method for Quantifying Viable Ascaris Eggs Using the First Internally Transcribed Spacer Region of Ribosomal DNA

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