+ Site Statistics
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

Evaluation of a PCR and culture assays to detect Aspergillus DNA in a murine model of invasive aspergillosis



Evaluation of a PCR and culture assays to detect Aspergillus DNA in a murine model of invasive aspergillosis



Blood 98(11 Part 1): 388a-389a



Invasive aspergillosis (IA) is one of the most common and life-threatening infections in patients receiving antileukemic therapy. Culture assays from blood samples are hardly ever positive even in pts. with IA. Preliminary results of PCR testing of blood samples in patients following intensive chemotherapy are rather promising To compare PCR and culture assays in detecting Aspergillosis spp. not only in blood specimens, but also in the target organ of invasive Aspergillosis, the lung, PCR and culture assay were evaluated in a murine model of invasive aspergillosis. Quantification of fungal load in blood and tissue was performed by quantitative culture assay and a quantitative Real-Time PCR assay established on the LightCycler. 82 mice were infected intranasally with Aspergillus conidia. Animals were sacrificed and defined sizes of lung tissue were homogenized and divided into 2 identical aliquots. 297/397 lung biopsy specimens (78%) showed a positive PCR result, whereas only 235/379 (62%) were positive by culture assay. Fungal load, as determined in 30 lung biopsy specimens by quantitative PCR assay, was 103-105 CFU/g. Sixty-four culture-negative lung biopsy specimens were positive by PCR, but only 2 PCR-negative samples were tested positive by culture assay. In order to compare the results of both assays, a statistical model was fitted in which the probability of a positive PCR result was calculated as a function of the amount of A. fumigatus-CFU. These results revealed the PCR assay to be 3.87 times more sensitive than the detection of fungi by culture for demonstrating fungal presence in lung biopsies (95%-CI: 2.96-5.01, p<0.0001). Blood specimens from 60 of these 82 mice infected intranasally with Aspergillus conidia were analyzed. 15/60 blood samples were PCR positive in contrast to none of the specimens tested positive by culture assay. When blood specimens were analyzed by quantitative PCR, fungal load was significantly lower when compared to lung biopsy specimens (101-102 vs 103-105 CFU/g). When lung biopsy specimens obtained from mice with PCR-negative blood specimens were compared to those with a positive PCR assay, median fungal load was significantly higher among the animals with a positive testing in blood samples (6500 vs 430 CFU/g). In conclusion, a superior sensitivity of the PCR assay when compared to culture could be confirmed in this murine model of invasive Aspergillosis. In addition, detection of Aspergillus-DNA in blood specimens correlated with a significantly higher fungal load in the lung further supporting screening blood samples in patients at risk of invasive Aspergillosis.

Please choose payment method:






(PDF emailed within 1 workday: $29.90)

Accession: 034878913

Download citation: RISBibTeXText


Related references

Detection of Aspergillus DNA by a nested PCR assay is superior to blood culture in an experimental murine model of invasive aspergillosis. Journal of Medical Microbiology 53(Pt 8): 803-806, 2004

Development of a monoclonal elisa to detect aspergillus antigenemia and clinical evaluation for diagnosis of invasive aspergillosis. Abstracts of the Annual Meeting of the American Society for Microbiology 87: 380, 1987

Improving the diagnosis of invasive aspergillosis by the detection of Aspergillus in broncho-alveolar lavage fluid: Comparison of non-culture-based assays. Journal of Infection 76(2): 196-205, 2018

Variation in virulence of Aspergillus fumigatus strains in a murine model of invasive pulmonary aspergillosis. Journal of Medical Microbiology 45(3): 186-191, 1996

Murine model of invasive pulmonary aspergillosis following an earlier stage, noninvasive Aspergillus infection. Journal of Clinical Microbiology 34(6): 1433-1439, 1996

Efficacy of LY303366 against amphotericin B-susceptible and -resistant Aspergillus fumigatus in a murine model of invasive aspergillosis. Antimicrobial Agents and ChemoTherapy 42(4): 873-878, 1998

The application of laser microdissection in molecular detection and identification of aspergillus fumigatus from murine model of acute invasive pulmonary aspergillosis. Mycopathologia 178(1-2): 53-61, 2014

RT-qPCR detection of Aspergillus fumigatus RNA in vitro and in a murine model of invasive aspergillosis utilizing the PAXgene® and Tempus™ RNA stabilization systems. Medical Mycology 50(6): 661-666, 2012

Isavuconazole (BAL4815) pharmacodynamic target determination in an in vivo murine model of invasive pulmonary aspergillosis against wild-type and cyp51 mutant isolates of Aspergillus fumigatus. Antimicrobial Agents and ChemoTherapy 57(12): 6284-6289, 2013

Evaluation of the efficacy of a posaconazole and anidulafungin combination in a murine model of pulmonary aspergillosis due to infection with Aspergillus fumigatus. Diagnostic Microbiology and Infectious Disease 90(1): 40-43, 2018

Efficacy of SCH-56592 in a temporarily neutropenic murine model of invasive aspergillosis with an itraconazole-susceptible and an itraconazole-resistant isolate of Aspergillus fumigatus. Antimicrobial Agents and ChemoTherapy 41(7): 1504-1507, 1997

Baseline serum Aspergillus galactomannan index in patients with hematologic malignancy and culture-documented invasive pulmonary aspergillosis: is there a difference among Aspergillus species?. Medical Mycology 2018:, 2018

Use of immunoblotting to detect Aspergillus fumigatus antigen in sera and urines of rats with experimental invasive aspergillosis. Journal of Clinical Microbiology 28(7): 1575-1579, 1990

Failure to detect circulating Aspergillus markers in a patient with chronic granulomatous disease and invasive aspergillosis. Journal of Clinical Microbiology 38(10): 3900-3901, 2000

Interstrain variability in the virulence of Aspergillus fumigatus and Aspergillus terreus in a Toll-deficient Drosophila fly model of invasive aspergillosis. Medical Mycology 48(2): 310-317, 2010