Related Experiment Video

Updated: Jul 19, 2026

Isolation, Characterization, and Total DNA Extraction to Identify Endophytic Fungi in Mycoheterotrophic Plants
06:53

Isolation, Characterization, and Total DNA Extraction to Identify Endophytic Fungi in Mycoheterotrophic Plants

Published on: May 5, 2023

A simple polymerase chain reaction-sequencing analysis capable of identifying multiple medically relevant filamentous

Timothy R Dean1, Michael Kohan, Doris Betancourt

  • 1National Risk Management Research Laboratory, U.S. Environmental Protection Agency, 109 T.W. Alexander Drive, Research Triangle Park, NC 27711, USA. dean.timothy@epa.gov

Mycopathologia
|October 14, 2006
PubMed

Insights

Accurate identification of indoor mold is crucial for health. A new PAN-PCR method combined with sequencing offers a rapid, specific, and cost-effective way to identify multiple fungal species, improving indoor air quality assessments.

Area of Science:

  • Environmental Science
  • Microbiology
  • Molecular Biology

Background:

  • Indoor environmental health issues are often linked to abnormal filamentous fungi (mold) growth.
  • Traditional mold identification relies on time-consuming and potentially inaccurate morphological assessments.
  • There is a need for rapid, sensitive, and precise methods for mold identification.

Purpose of the Study:

  • To develop and validate a novel method for the unambiguous identification of multiple fungal organisms.
  • To provide a rapid, specific, and cost-effective alternative to traditional mold identification techniques.

Main Methods:

  • A PAN-PCR (Polymerase Chain Reaction) approach was employed using universal primers to amplify ribosomal DNA sequences.
  • Amplified DNA sequences were cloned into Escherichia coli.
  • Individual clones were sequenced and analyzed for unambiguous identification of fungal species.

Main Results:

  • The PAN-PCR method successfully identified multiple fungal species, including Stachybotrys chartarum, Penicillium purpurogenum, Aspergillus sydowii, and Cladosporium cladosporioides from a mixed culture.
  • The method demonstrated high specificity and accuracy.
  • The protocol was found to be rapid, easy to perform, and cost-effective.

Conclusions:

  • The devised PAN-PCR coupled with cloning and sequencing is a powerful tool for accurate and timely identification of indoor mold species.
  • This method addresses the limitations of traditional morphological identification, offering significant advantages for indoor environmental quality assessments.
  • The approach facilitates precise identification of fungi, contributing to better understanding and management of health-related issues linked to indoor mold exposure.

More Related Videos

Whole Genome Sequencing of Candida glabrata for Detection of Markers of Antifungal Drug Resistance08:45

Whole Genome Sequencing of Candida glabrata for Detection of Markers of Antifungal Drug Resistance

Published on: December 28, 2017

Related Concept Videos

Modern Molecular Taxonomy01:29

Modern Molecular Taxonomy

Advancements in molecular biology have revolutionized the identification and characterization of bacteria, with multiple methods leveraging DNA sequencing for enhanced precision. As sequencing technologies improve and costs decline, these approaches are increasingly used in clinical, environmental, and evolutionary studies.Multilocus Sequence Typing (MLST) examines several housekeeping genes, essential chromosomal genes encoding cellular functions, to distinguish strains. Approximately...
Rapid Identification of Pathogens01:25

Rapid Identification of Pathogens

MALDI-TOF MS has transformed clinical microbiology by offering a rapid and reliable method for pathogen identification. The traditional approach to microbial identification typically involves time-consuming culture techniques and biochemical tests, which can delay the initiation of appropriate antimicrobial therapy. MALDI-TOF MS avoids these delays by using characteristic ribosomal protein mass patterns of microbial cells, enabling accurate species-level identification within minutes.Principle...
Sanger Sequencing01:57

Sanger Sequencing

DNA sequencing is a fundamental technique that is routinely used in the biological sciences. This method can be applied to a range of questions at different scales - from the sequencing of a cloned DNA fragment or the study of a mutation in a gene up to whole-genome sequencing. However, despite the widespread use of sequencing today, it was not until 1977 that Fredrick Sanger and his collaborators developed the chain-termination method to decode DNA sequences. It relies on the separation of a...
RNA-seq03:21

RNA-seq

RNA sequencing, or RNA-Seq, is a high-throughput sequencing technology used to study the transcriptome of a cell. Transcriptomics helps to interpret the functional elements of a genome and identify the molecular constituents of an organism. Additionally, it also helps in understanding the development of an organism and the occurrence of diseases. 
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while microarray-based...
Applications of Molecular Taxonomy01:20

Applications of Molecular Taxonomy

Molecular taxonomy has revolutionized the understanding and classification of bacteria, providing precise insights into their diversity, evolutionary relationships, and ecological roles. By utilizing molecular techniques such as DNA sequencing and fingerprinting, researchers have made significant strides in various fields related to bacterial studies.Resolving Taxonomic AmbiguitiesMolecular taxonomy has been instrumental in distinguishing closely related bacterial species initially thought to...
Next-generation Sequencing03:00

Next-generation Sequencing

The first human genome sequencing project cost $2.7 billion and was declared complete in 2003, after 15 years of international cooperation and collaboration between several research teams and funding agencies. Today, with the advent of next-generation sequencing technologies, the cost and time of sequencing a human genome have dropped over 100 fold.
Next-Generation Sequencing Methods
Although all next-generation methods use different technologies, they all share a set of standard features.