Detection of Pneumocystis jirovecii in oral wash from immunosuppressed patients as a diagnostic tool

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 ONE

JF - PLOS ONE

IS - 3

M1 - e0174012

ER -