TY - JOUR
T1 - Detection of Pneumocystis jirovecii in oral wash from immunosuppressed patients as a diagnostic tool
AU - Hviid, Cecilie Juul
AU - Ellermann-Eriksen, Svend
AU - Jespersen, Bente
AU - Dam, Mette Yde
AU - Dahlerup, Jens Frederik
AU - Benfield, Thomas
AU - Jespersen, Sanne
AU - Østergaard, Lars Jørgen
AU - Laursen, Alex Lund
PY - 2017/3/1
Y1 - 2017/3/1
N2 - BACKGROUND: Diagnosis of Pneumocystis jirovecii (PJ) pneumonia ordinarily requires invasive procedures that could be avoided by PCR methodologies, if these could be designed with adequate cut-off values for confounding background carriage.METHODS: We designed a novel quantitative real-time PCR assay to detect the mitochondrial large subunit rRNA gene of PJ in oral washes. To benchmark levels of PJ carriage versus infection, we tested asymptomatic immunosuppressed patients including Danish (n = 88) and West African HIV-infected (n = 142) patients, renal transplant recipients (n = 51), rheumatologic patients (n = 102), patients with inflammatory bowel diseases (n = 98), and healthy blood donors (controls, n = 50). The fungal burden in patients with PJ pneumonia (PCP, n = 7) was also investigated.RESULTS: Danish HIV-infected patients (with viremia/low CD4) and recent transplant recipients were at most risk of being carriers (prevalence of 23% and 16.7% respectively), whereas PJ was rarely detected among rheumatologic patients, patients with inflammatory bowel diseases, and untreated West African HIV patients. PJ was not detected among healthy controls. The fungal burden in patients with PCP fell rapidly on treatment.CONCLUSIONS: The quantitative PCR method described could conceivably discriminate between carriage and disease, given suitable threshold values for the former, and predict treatment efficacy by measures of the fungal burden in daily oral washes.
AB - BACKGROUND: Diagnosis of Pneumocystis jirovecii (PJ) pneumonia ordinarily requires invasive procedures that could be avoided by PCR methodologies, if these could be designed with adequate cut-off values for confounding background carriage.METHODS: We designed a novel quantitative real-time PCR assay to detect the mitochondrial large subunit rRNA gene of PJ in oral washes. To benchmark levels of PJ carriage versus infection, we tested asymptomatic immunosuppressed patients including Danish (n = 88) and West African HIV-infected (n = 142) patients, renal transplant recipients (n = 51), rheumatologic patients (n = 102), patients with inflammatory bowel diseases (n = 98), and healthy blood donors (controls, n = 50). The fungal burden in patients with PJ pneumonia (PCP, n = 7) was also investigated.RESULTS: Danish HIV-infected patients (with viremia/low CD4) and recent transplant recipients were at most risk of being carriers (prevalence of 23% and 16.7% respectively), whereas PJ was rarely detected among rheumatologic patients, patients with inflammatory bowel diseases, and untreated West African HIV patients. PJ was not detected among healthy controls. The fungal burden in patients with PCP fell rapidly on treatment.CONCLUSIONS: The quantitative PCR method described could conceivably discriminate between carriage and disease, given suitable threshold values for the former, and predict treatment efficacy by measures of the fungal burden in daily oral washes.
KW - Adult
KW - Africa, Western
KW - DNA, Fungal/genetics
KW - Denmark
KW - HIV Infections/complications
KW - Humans
KW - Immunocompromised Host/genetics
KW - Kidney Transplantation/adverse effects
KW - Male
KW - Middle Aged
KW - Pneumocystis carinii/genetics
KW - Pneumonia, Pneumocystis/complications
U2 - 10.1371/journal.pone.0174012
DO - 10.1371/journal.pone.0174012
M3 - Journal article
C2 - 28358900
SN - 1932-6203
VL - 12
JO - PLoS Computational Biology
JF - PLoS Computational Biology
IS - 3
M1 - e0174012
ER -
