Optimization of a real-time PCR assay to quantitate airborne fungi collected on a gelatin filter

Journal of Bioscience and Bioengineering
Naomichi YamamotoYukio Yanagisawa

Abstract

The present study aimed to optimize a real-time PCR assay to quantitate airborne fungi collected on a gelatin filter. In particular, the study optimized conditions for the DNA extraction and real-time PCR amplification to accurately measure airborne fungal concentrations. First, time of fine bead homogenization to extract the DNA from fungal cells was optimized to maximize the DNA yield and prepare the DNA suitable for sensitive and precise quantification by a subsequent real-time PCR analysis. Second, a condition for the real-time PCR amplification was optimized to successfully amplify and quantitate the extracted fungal DNA. In particular, a dilution ratio of the DNA extracts to be introduced to PCR was optimized to achieve an appropriate balance between mitigating PCR inhibition and securing detection sensitivity. Since concentrations of airborne fungi generally observed in indoor and outdoor environments (i.e., 10(1)-10(4) CFU m(-3)) were found to be near the limit of quantification by the generally-used molecular-based detection technique in conjunction with use of gelatin filters, optimizations of these conditions were found to be crucial. Our preliminary result showed that a culture-based method underestimated concentrat...Continue Reading

References

Jan 1, 1988·The Annals of Occupational Hygiene·W Eduard, O Aalen
Jan 1, 1988·The Annals of Occupational Hygiene·P HeikkiläK Louhelainen
Feb 1, 1984·American Industrial Hygiene Association Journal·J M Macher, M W First
Feb 15, 2001·Clinical Infectious Diseases : an Official Publication of the Infectious Diseases Society of America·S J LinS M Teutsch
May 26, 2001·Applied and Environmental Microbiology·R H WilliamsH A McCartney
Jul 4, 2002·Journal of Environmental Monitoring : JEM·Zhihong WuGöran Blomquist
Nov 6, 2004·Applied and Environmental Microbiology·W Elliott HornerPhilip R Morey
Dec 7, 2005·Environmental Health Perspectives·Robert WålinderGunilla Wieslander
Jan 6, 2006·Journal of Environmental Monitoring : JEM·Qing-Yin ZengXiao-Ru Wang
Aug 3, 2006·Journal of Forensic Sciences·Elias J Kontanis, Floyd A Reed
Jun 5, 2007·Indoor Air·D Mudarri, W J Fisk
Apr 1, 2009·Journal of Bioscience and Bioengineering·Naomichi YamamotoYukio Yanagisawa

❮ Previous
Next ❯

Citations

Sep 8, 2010·Applied and Environmental Microbiology·Denina HospodskyJordan Peccia
Apr 17, 2013·Critical Reviews in Microbiology·Delphine MéheustJean-Pierre Gangneux
Feb 9, 2019·Molecular Ecology Resources·Robert M W FergusonIan Colbeck

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.