Screening for PTLD in lung and heart–lung transplant recipients by measuring EBV DNA load in bronchoalveolar lavage fluid using real time PCR

Abstract

Pediatric L-HLTx recipients are at risk for developing PTLD with the lung being a primary site of disease. We hypothesized that BALF is a better sample than peripheral blood for measuring EBV DNA load in this high-risk population. Archived BALF specimens from pediatric L-HLTx recipients with and without PTLD were assayed for EBV DNA load using a quantitative real time TaqMan PCR assay. These values were compared with values determined in peripheral blood by a competitive PCR assay. Fifty-five BALF specimens from 16 L-HLTx patients were evaluated. Three patients with PTLD had mean BALF EBV DNA load values almost 50-fold higher than subjects without PTLD (4.6 x 10(5) copies/mL vs. 1.0 x 10(4) copies/mL). Patients who were EBV seronegative pretransplantation (i.e., high risk for PTLD) had elevated EBV DNA load values vs. patients who were EBV seropositive pretransplantation, regardless of the diagnosis of PTLD (mean values of 3.2 x 10(5) copies/mL vs. 1.1 x 10(4) copies/mL). Lastly, BALF analysis identified all subjects with PTLD, whereas peripheral blood analysis identified only one of these cases. Therefore, it can be concluded that monitoring EBV DNA load in BALF following L-HLTx facilitates detection of PTLD in high-risk patients and may be superior to peripheral blood assays.

Loads, lungs, and lymphoproliferative disorders: role of Epstein-Barr virus and limitations of what we know

“…Had the clinicians caring for the patient based their diagnosis only on the EBV viral load in the blood, this diagnosis might have been missed. The finding of a positive EBV load in BAL fluid in lung transplant recipients with low or absent EBV load in blood, as was seen in this case, has been previously reported by Michelson et al (8). The study looked at 54 stored BAL samples from 16 pediatric lung transplant recipients and generally found negative to low levels of EBV in patients without lung disease but high levels of EBV in the BAL of 3 children who had PTLD.…”

Acute cellular rejection and Epstein-Barr virus-related post-transplant lymphoproliferative disorder in a pediatric lung transplant with low viral load

“…A variety of factors may influence viral shedding of EBV by the tumor (e.g., the presence of a lytic vs. latent infection, or the ability of the patient's immune response to partially control viral shedding) (14). BAL, which is routinely performed as a part of post‐transplant surveillance, has been demonstrated as a more sensitive diagnostic tool for the detection of EBV DNA in thoracic transplant recipients (22), as in our case. Morphological and molecular evaluation remains the gold standard for precise characterization of PTLD.…”

Posttransplant Lymphoproliferative Disease after Lung Transplantation

“…Michelson and coworkers analyzed bronchoalveolar lavage (BAL) fluid from 16 heart-lung transplant patients for quantitative PCR for EBV and found values 50 times higher in patients with PTLD. BAL proved to be highly sensitive in detecting 3/3 cases of PTLD compared to testing of peripheral blood [38]. …”