The use of real-time PCR and species-specific primers for the identification and monitoring of Paecilomyces lilacinus

• Published in: FEMS microbiology ecology Vol. 51; no. 2; pp. 257 - 264
• Main Authors: Atkins, Simon D., Clark, Ian M., Pande, Sonal, Hirsch, Penny R., Kerry, Brian R.
• Format: Journal Article
• Language: English
• Published: Oxford, UK Elsevier B.V 2005; Blackwell Publishing Ltd; Blackwell; Oxford University Press
• Subjects: Animals; Baits; Biological and medical sciences; Clinical isolates; DNA Primers - genetics; Ecology; Eggs; Fundamental and applied biological sciences. Psychology; Fungi; Medical sciences; Microbiology; Miscellaneous; Mycology; Nematoda - microbiology; Nematodes; Ovum - microbiology; Paecilomyces - classification; Paecilomyces - genetics; Paecilomyces - isolation & purification; Paecilomyces lilacinus; PCR diagnostics; Polymerase Chain Reaction - methods; Population number; Potato cyst nematodes; Potatoes; Primers; Real time; Real-time PCR; Soil Microbiology; Species Specificity; Species-specific primers; Tumors; Paecilomyces lilacinus; Species-specific primers; PCR diagnostics; Real-time PCR; Potato cyst nematodes; Species specificity; Fungi; Paecilomyces; Gene; Dicotyledones; Angiospermae; Cyst; Fungi Imperfecti; Solanaceae; Benign neoplasm; Diagnosis; Clinical isolate; Nematoda; Thallophyta; Human; Root; Nucleotide sequence; Solanum tuberosum; Real time; Polymerase chain reaction; Soils; Treatment; Helmintha; Nemathelminthia; Spermatophyta; Specific identification; Tuber plant; Invertebrata; Detection
• ISSN: 0168-6496; 1574-6941
• DOI: 10.1016/j.femsec.2004.09.002

The
Paecilomyces lilacinus is the most widely tested fungus for the control of root-knot and cyst nematodes. The fungus has also been implicated in a number of human and animal infections, difficulties in diagnosis often result in misdiagnosis or delays in identification leading to a delay in treatment. Here, we report the development of species-specific primers for the identification of
P. lilacinus based on sequence information from the ITS gene, and their use in identifying
P. lilacinus isolates, including clinical isolates of the fungus. The primer set generated a single PCR fragment of 130 bp in length that was specific to
P. lilacinus and was also used to detect the presence of
P. lilacinus from soil, roots and nematode eggs. Real-time PCR primers and a TaqMan
® probe were also developed and provided quantitative data on the population size of the fungus in two field sites. PCR, bait and culture methods were combined to investigate the presence and abundance of the fungus from two field sites in the United Kingdom where potato cyst nematode populations were naturally declining, and results demonstrated the importance of using a combination of methods to investigate population size and activity of fungi.