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Published on: February 1, 2014
Application of nested PCR to detect Penicillium marneffei in serum samples
Monsicha Pongpom1, Thira Sirisanthana, Nongnuch Vanittanakom
1Department of Microbiology, Chiang Mai University, Chiang Mai, Thailand.
Abstract:
We previously reported a nested PCR assay for specific identification of 18S ribosomal DNA of Penicillium marneffei. In this study, the assay was used to detect the DNA of P. marneffei in serum samples. Sensitivity of the test was 4 pg/microl and 0.4 fg/microl when the cycle numbers used for nested reactions were 15 and 30, respectively. Twenty four out of 35 sera (68.6%) collected from patients with culture confirmed penicilliosis marneffei were positive, while normal healthy and non-P. marneffei infected HIV-positive sera were negative. The results suggested that the assay could be applied for the diagnosis of infections due to P. marneffei.
Insights
A novel nested PCR assay accurately detects Penicillium marneffei DNA in patient serum samples. This diagnostic tool shows high sensitivity, aiding in the diagnosis of penicilliosis marneffei infections.
Area of Science:
- Medical Mycology
- Molecular Diagnostics
- Infectious Diseases
Background:
- Penicillium marneffei is an emerging fungal pathogen causing systemic infections, particularly in immunocompromised individuals.
- Accurate and rapid diagnostic methods are crucial for timely treatment of penicilliosis marneffei.
Purpose of the Study:
- To evaluate a previously developed nested PCR assay for the detection of Penicillium marneffei DNA in clinical serum samples.
- To determine the sensitivity and specificity of the assay for diagnosing penicilliosis marneffei.
Main Methods:
- A nested PCR assay targeting the 18S ribosomal DNA of Penicillium marneffei was employed.
- The assay's sensitivity was assessed using known concentrations of P. marneffei DNA.
- Serum samples from patients with confirmed penicilliosis marneffei and control groups were tested.
Main Results:
- The nested PCR assay demonstrated high sensitivity, detecting as low as 4 pg/microl (15 cycles) and 0.4 fg/microl (30 cycles) of P. marneffei DNA.
- 68.6% (24/35) of serum samples from patients with culture-confirmed penicilliosis marneffei were positive.
- All sera from healthy individuals and HIV-positive patients without P. marneffei infection tested negative.
Conclusions:
- The nested PCR assay is a sensitive and specific method for detecting Penicillium marneffei DNA in serum.
- This molecular assay holds significant potential for the clinical diagnosis of penicilliosis marneffei infections.
