Detection and Identification of Toxic Microalgae by the Use of Innovative Molecular Methods

SUMMARYThe development of simple, sensitive and rapid methods for the detection and identification ofToxoplasma gondiiis important for the diagnosis and epidemiological studies of the zoonotic disease toxoplasmosis. In the past 2 decades, molecular methods based on a variety of genetic markers have been developed, each with its advantages and limitations. The application of these methods has generated invaluable information to enhance our understanding of the epidemiology, population genetics and phylogeny ofT. gondii. However, since most studies focused solely on the detection but not genetic characterization ofT. gondii, the information obtained was limited. In this review, we discuss some widely used molecular methods and propose an integrated approach for the detection and identification ofT. gondii, in order to generate maximum information for epidemiological, population and phylogenetic studies of this key pathogen.

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Molecular Detection and Identification of Brettanomyces/Dekkera bruxellensis and Brettanomyces/Dekkera anomalus in Spoiled Wines

Applied and Environmental Microbiology ◽

10.1128/aem.70.3.1347-1355.2004 ◽

2004 ◽

Vol 70 (3) ◽

pp. 1347-1355 ◽

Cited By ~ 65

Author(s):

Luca Cocolin ◽

Kalliopi Rantsiou ◽

Lucilla Iacumin ◽

Roberto Zironi ◽

Giuseppe Comi

Keyword(s):

Direct Detection ◽

Molecular Methods ◽

Enzyme Analysis ◽

Restriction Enzyme Analysis ◽

Rrna Gene ◽

Brettanomyces Bruxellensis ◽

Dna And Rna ◽

Detection And Identification ◽

26S Rrna ◽

Total Agreement

ABSTRACT In this paper we describe the development of a PCR protocol to specifically detect Brettanomyces bruxellensis and B. anomalus. Primers DB90F and DB394R, targeting the D1-D2 loop of the 26S rRNA gene, were able to produce amplicons only when the DNA from these two species were used. No amplification product was obtained when DNA from other Brettanomyces spp. or wine yeasts were used as the templates. The 305-bp product was subjected to restriction enzyme analysis with DdeI to differentiate between B. bruxellensis and B. anomalus, and each species could be identified on the basis of the different restriction profiles. After optimization of the method by using strains from international collections, wine isolates were tested with the method proposed. Total agreement between traditional identification and molecular identification was observed. The protocol developed was also used for direct detection of B. bruxellensis and B. anomalus in wines suspected to be spoiled by Brettanomyces spp. Application of culture-based and molecular methods led us to the conclusion that 8 of 12 samples were spoiled by B. bruxellensis. Results based on the application of molecular methods suggested that two of the eight positive samples had been infected more recently, since specific signals were obtained at both the DNA and RNA levels.

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An Overview of the Molecular Methods in the Diagnosis of Gastrointestinal Infectious Diseases

International Journal of Microbiology ◽

10.1155/2020/8135724 ◽

2020 ◽

Vol 2020 ◽

pp. 1-13

Author(s):

Muhammad Amjad

Keyword(s):

Infectious Diseases ◽

Diagnostic Techniques ◽

Antigen Detection ◽

Molecular Methods ◽

Advantages And Disadvantages ◽

Laboratory Setup ◽

Detection And Identification ◽

Intestinal Infections ◽

Wide Range ◽

Trained Staff

Gastrointestinal infectious diseases are very common worldwide and an important cause of morbidity and mortality, particularly in infants in developing countries. Diarrhea and other intestinal infections are caused by a wide range of bacteria, viruses, protozoa, and parasites. Conventional diagnosis of these infections is performed by culture, microscopy, and antigen detection immunoassays. The traditional culture and microscopy procedures are time-consuming, lack sensitivity, and require special laboratory setup and well-trained staff. However, based on the advancement in the molecular diagnostics and with the introduction of commercially available tests, traditional diagnostic techniques have been continuously replaced by these newer rapid antigen detection and molecular-based methods. This review summarizes and discusses the availability, advantages, and disadvantages of molecular methods in the detection and identification of human gastrointestinal pathogens.

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Use of Molecular Methods for the Detection and Identification of Wood Decay Fungi

Forest Products Biotechnology ◽

10.1201/9781482272734-21 ◽

1997 ◽

pp. 315-330

Keyword(s):

Wood Decay ◽

Molecular Methods ◽

Decay Fungi ◽

Detection And Identification ◽

Wood Decay Fungi

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Molecular detection and identification of 'Candidatus Liberibacter solanacearum'

Biljni lekar ◽

10.5937/biljlek2101017m ◽

2021 ◽

Vol 49 (1) ◽

pp. 17-37

Author(s):

Jelena Menković ◽

Aleksa Obradović

Keyword(s):

Host Plants ◽

Molecular Methods ◽

Cultivated Plants ◽

Economic Losses ◽

Insect Vectors ◽

Long Distance ◽

Candidatus Liberibacter Solanacearum ◽

Detection And Identification ◽

Geographical Regions ◽

Candidatus Liberibacter

"Candidatus Liberibacter solanacearum" is a gram-negative, fastidious bacterium that inhabits and multiplies in the phloem of host plants. The bacterium causes economically important diseases of solanaceous crops such as "zebra chips" of potatoes, and chlorotic weakening of cultivated plants of the Apiaceae family. Beside potatotes, this pathogen causes significant economic losses in tobacco, peper and tomato production. Insect vectors classified in the family Triozidae play an important role in the epidemiology of this bacterium. Insects transfer bacterial cells from infected to healthy plants by feeding on the plant juice from the phloem tissue. This pathogenic bacterium could be transmitted by infected seeds which result in long-distance spread. Based on the analysis of single nucleotide polymorphism (SNP), nine haplotypes (A, B, C, D, E, F, G, H, U) from different geographical regions, host plants and insect vectors have been described. Considering to impossibility of isolation of this bacterium on a nutrient medium, detection and identification are based on the application of molecular methods (conventional PCR, real-time PCR, multiplex PCR, nested PCR and LAMP PCR). Taking into consideration the increasing spread of "Candidatus Liberibacter solanacearum" in Europe, as well as its recent detection in Serbia, the aim of this paper was to raise an awareness about importance of this bacterium and summarize the protocols and molecular methods for its detection and identification.

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