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. 2001 Apr;39(4):1506-9.
doi: 10.1128/JCM.39.4.1506-1509.2001.

Diagnosis and monitoring of murine histoplasmosis by a nested PCR assay

Affiliations

Diagnosis and monitoring of murine histoplasmosis by a nested PCR assay

R Bialek et al. J Clin Microbiol. 2001 Apr.

Abstract

A newly developed nested PCR assay was applied to murine models of histoplasmosis. ICR and BALB/c mice were intravenously infected with Histoplasma capsulatum and sacrificed up to 29 days later. Samples of blood, spleen, and lung homogenates were cultured and examined by the PCR assay. In the ICR mouse model, 265 of 319 organ samples showed concordant results. With 7 samples, the culture was positive and the PCR assay was negative whereas a positive PCR but a negative culture were obtained with 47 samples (P < 0.0001 according to McNemar's test). Organ homogenates and blood samples of either spontaneously cured or treated BALB/c mice were PCR negative. The nested PCR assay performs excellently in the monitoring of spontaneously and treatment-cured murine histoplasmosis. It limits the infection risks of the laboratory staff and might be of diagnostic value for humans.

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Figures

FIG. 1
FIG. 1
H. capsulatum tissue burdens of lungs and spleens detected by PCR and culture. On each day p.i., 64 samples, except on day 5 p.i. (only 63 samples) from eight ICR mice were examined.
FIG. 2
FIG. 2
(a) Probability of a positive H. capsulatum culture result, depending on the fungal concentration in spleen tissue of infected ICR mice. Vertical bars, 95% CI and observed proportion of a positive culture result; curve, expected probability of a positive culture. Point of inflection log concentration = −0.49 (95% CI, −0.71 to −0.27); slope parameter, 3.0 (95% CI, 2.0 to 4.4). (b) Probability of a positive H. capsulatum nested PCR assay result, depending on the fungal concentration in spleen tissue of infected ICR mice. Vertical bars, 95% CI and observed proportion of a positive nested PCR assay result; curve, expected probability of a positive nested PCR assay result. Point of inflection log concentration = −0.57 (95% CI, −0.88 to −0.27); slope parameter, 2.2 (95% CI, 1.4 to 3.5); asymptote for very low concentrations, 0.125 (95% CI, 0.066 to 0.201).

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