Rapid authentication of the precious herb saffron by loop-mediated isothermal amplification (LAMP) based on internal transcribed spacer 2 (ITS2) sequence

Abstract Background The malaria mosquito Anopheles punctipennis, a widely distributed species in North America, is capable of transmitting human malaria and is actively involved in the transmission of the ungulate malaria parasite Plasmodium odocoilei. However, molecular diagnostic tools based on Internal Transcribed Spacer 2 (ITS2) of ribosomal DNA are lacking for this species. Anopheles punctipennis is a former member of the Anopheles maculipennis complex but its systematic position remains unclear. Methods In this study, ITS2 sequences were obtained from 276 An. punctipennis specimens collected in the eastern and midwestern United States and a simple and robust Restriction Fragment Length Polymorphism approach for species identification was developed. The maximum-likelihood phylogenetic tree was constructed based on ITS2 sequences available through this study and from GenBank for 20 species of Anopheles. Results The analysis demonstrated a consistent ITS2 sequence length and showed no indications of intragenomic variation among the samples based on ITS2, suggesting that An. punctipennis represents a single species in the studied geographic locations. In this study, An. punctipennis was found in urban, rural, and forest settings, suggesting its potential broad role in pathogen transmission. Phylogeny based on ITS2 sequence comparison demonstrated the close relationship of this species with other members of the Maculipennis group. Conclusions This study developed molecular tools based on ITS2 sequences for the malaria vector An. punctipennis and clarified the phylogenetic position of the species within the Maculipennis group.

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Loop-mediated Isothermal Amplification and nested PCR of the Internal Transcribed Spacer (ITS) for Histoplasma capsulatum detection

PLoS Neglected Tropical Diseases ◽

10.1371/journal.pntd.0007692 ◽

2019 ◽

Vol 13 (8) ◽

pp. e0007692 ◽

Cited By ~ 4

Author(s):

Matheus da Silva Zatti ◽

Thales Domingos Arantes ◽

José Alex Lourenço Fernandes ◽

Mônica Baumgardt Bay ◽

Eveline Pipolo Milan ◽

...

Keyword(s):

Nested Pcr ◽

Internal Transcribed Spacer ◽

Histoplasma Capsulatum ◽

Isothermal Amplification ◽

Loop Mediated Isothermal Amplification

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Evolution and function of B chromosome 45S rDNA sequences in Brachycome dichromosomatica

Genome ◽

10.1139/g07-048 ◽

2007 ◽

Vol 50 (7) ◽

pp. 638-644 ◽

Cited By ~ 15

Author(s):

Sylvia Marschner ◽

Armin Meister ◽

Frank R. Blattner ◽

Andreas Houben

Keyword(s):

Phylogenetic Analysis ◽

Internal Transcribed Spacer ◽

B Chromosome ◽

Its2 Sequence ◽

B Chromosomes ◽

Rrna Gene ◽

45S Rdna ◽

Internal Transcribed Spacer 2 ◽

Rdna Sequences ◽

And Function

The origin and activity of 45S rDNA located on micro B chromosomes of the daisy Brachycome dichromosomatica were analysed. The internal transcribed spacer 2 (ITS2) of the 45S rRNA gene was sequenced for micro B, large B, and A chromosomes of B. dichromosomatica cytodeme A2, and conserved differences were identified between sequences originating from A and both types of B chromosomes. Phylogenetic analysis did not identify a species containing an ITS2 sequence more similar to either of the B chromosome sequences than the B. dichromosomatica A chromosome sequences. Thus, an origin of the B chromosomes from A chromosomes at a time prior to the divergence of the 4 cytodemes of B. dichromosomatica is suggested. The frequent (70%) nucleolar non-association of micro B chromosomes suggests inactivity of micro B 45S rDNA.

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Placozoa: at least two

Biologia ◽

10.2478/s11756-007-0143-z ◽

2007 ◽

Vol 62 (6) ◽

Cited By ~ 9

Author(s):

Matthias Wolf ◽

Christian Selig ◽

Tobias Müller ◽

Nicole Philippi ◽

Thomas Dandekar ◽

...

Keyword(s):

Secondary Structure ◽

Internal Transcribed Spacer ◽

Distinct Species ◽

Secondary Structures ◽

Its2 Sequence ◽

Structure Alignment ◽

Sequence Structure ◽

Rna Sequence ◽

Internal Transcribed Spacer 2 ◽

Compensatory Base Changes

AbstractIt was shown that compensatory base changes (CBCs) in internal transcribed spacer 2 (ITS2) sequence-structure alignments can be used for distinguishing species. Using the ITS2 Database in combination with 4SALE — a tool for synchronous RNA sequence and secondary structure alignment and editing — in this study we present an in-depth CBC analysis for placozoan ITS2 sequences and their respective secondary structures. This analysis indicates at least two distinct species in Trichoplax (Placozoa) supporting a recently suggested hypothesis, that Placozoa is “no longer a phylum of one”.

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Rapid identification of Hebeloma crustuliniforme species using real-time fluorescence and visual loop-mediated isothermal amplification based on the internal transcribed spacer sequence

Food Control ◽

10.1016/j.foodcont.2021.108301 ◽

2021 ◽

pp. 108301

Author(s):

Jie Gao ◽

Nan Wang ◽

Juan Zhang ◽

Ruibin Xie ◽

Wenjie Yu ◽

...

Keyword(s):

Real Time ◽

Internal Transcribed Spacer ◽

Rapid Identification ◽

Isothermal Amplification ◽

Loop Mediated Isothermal Amplification ◽

Hebeloma Crustuliniforme

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Rapid and reliable distinguish of Morinda officinalis How by loop-mediated isothermal amplification (LAMP) assay

10.21203/rs.3.rs-120446/v1 ◽

2020 ◽

Author(s):

Shuhan Wang ◽

Aihua Sui ◽

Yafei Han ◽

Quan Zhou ◽

Hang Xu ◽

...

Keyword(s):

Low Cost ◽

Lamp Assay ◽

Its2 Sequence ◽

Isothermal Amplification ◽

Nucleic Acid Amplification ◽

Nuclear Ribosomal Dna ◽

Traditional Medicines ◽

Loop Mediated Isothermal Amplification ◽

Amplification Method ◽

Human Disorders

Abstract The medicinal plant Morinda officinalis How (MO), especially the root, has been frequently used in traditional medicines around the world as an herbal drug for treating variable human disorders and diseases. Various adulterations of MO were found for economic or production limitations. However, authentication of MO from its adulterants by LAMP has not yet been established. The present study introduces a commercially available nucleic acid amplification method, loop-mediated isothermal amplification (LAMP) assay for the distinguish of MO from its adulterants. In this method, we combined DNA barcodes technology to design 2 pairs independent LAMP primers, which based on the internal transcribed spacer 2 (ITS2) sequence of MO’s nuclear ribosomal DNA. Our results showed that the LAMP could amplify the samples as expected and successfully identify target MO, and the limit for DNA template preciseness was verified as 1 × 10− 1 pg/µl. All the visual or real-time turbidity detection was performed within 60 min at approximately 63 °C. The result showed that the LAMP assay and primers we designed have high accuracy and efficiency for the differentiation of MO and its adulterants. Our results illustrated that the proposed low-cost, fast and reliable LAMP assay without the need for expensive equipment or specialized techniques could be a good way for MO rapid authentication.

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Orthopodomyia pulcripalpis (Diptera: Culicidae), a genus and species new to the Iranian mosquito fauna, with a review of bionomical information*

Zootaxa ◽

10.11646/zootaxa.4299.1.11 ◽

2017 ◽

Vol 4299 (1) ◽

pp. 141 ◽

Cited By ~ 4

Author(s):

SHAHYAD AZARI-HAMIDIAN ◽

BEHZAD NOROUZI ◽

AYOOB NOORALLAHI

Keyword(s):

New Species ◽

Aedes Albopictus ◽

Internal Transcribed Spacer ◽

Sequence Data ◽

Its2 Sequence ◽

Internal Transcribed Spacer 2 ◽

As Species ◽

Southwestern Iran ◽

A New Species

The most recent checklist of Iranian mosquitoes (Diptera: Culicidae) includes 64 species representing seven genera (Azari-Hamidian, 2007; Azari-Hamidian & Harbach, 2009). Subsequently, Oshaghi et al. (2008) found that Anopheles superpictus Grassi is two species in Iran based on the Internal Transcribed Spacer 2 (ITS2) sequences of rDNA, which were later listed as species A and B by Harbach (2013), and Djadid et al. (2009) recognized a new species of the Anopheles hyrcanus group (An. hyrcanus spIR) from southwestern Iran, also based on ITS2 sequence data. More recently, Doosti et al. (2016) reported the occurrence of Aedes albopictus (Skuse) in southeastern Iran.

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The New Internal Transcribed Spacer 2 Diagnostic Tool Clarifies the Taxonomic Position and Geographic Distribution of the North American Malaria Vector Anopheles Punctipennis

10.21203/rs.3.rs-83154/v1 ◽

2020 ◽

Author(s):

James M. Hodge ◽

Andrey A. Yurchenko ◽

Dmitriy A. Karagodin ◽

Reem A. Masri ◽

Ryan C. Smith ◽

...

Keyword(s):

Malaria Vector ◽

Internal Transcribed Spacer ◽

Single Species ◽

Its2 Sequence ◽

Pathogen Transmission ◽

Molecular Diagnostic ◽

Diagnostic Tools ◽

Sequence Length ◽

Phylogenetic Position ◽

Internal Transcribed Spacer 2

Abstract Background: The malaria mosquito Anopheles punctipennis Say, a widely distributed species in North America, is capable of transmitting human malaria and is actively involved in the transmission of the ungulate malaria parasite Plasmodium odocoilei. However, molecular diagnostic tools based on Internal Transcribed Spacer 2 (ITS2) of ribosomal DNA are lacking for An. punctipennis. An. punctipennis is a former member of the Anopheles maculipennis complex but its systematic position remains unclear. Results: In this study, we collected and sequenced ITS2 from 276 An. punctipennis specimens collected from the eastern and midwestern United States. Our analysis demonstrated a consistent sequence length and showed no indications of intragenomic variation among the samples based on ITS2, suggesting that An. punctipennis represents a single species in the studied geographic locations. In addition to the ITS2 sequence-based molecular tool, we developed a simple and robust Restriction Length Polymorphism approach for species identification. An. punctipennis was found in urban, rural, and forest settings, suggesting its potential broad role in pathogen transmission. Phylogeny based on ITS2 sequence comparison demonstrated the close relationship of this species with other members of the Maculipennis group. Conclusions: This study developed molecular tools based on ITS2 sequences for the malaria vector An. punctipennis and clarified the phylogenetic position of the species within the Maculipennis group.

Download Full-text