Multiplexed PCR genotyping of HPVs from plantaris verrucae.

BACKGROUND: Plantaris verrucae are a common diagnosis in childhood, consume a significant amount of health-care resources, have many painful treatment options and many recurrences. OBJECTIVES: The objective of this study was to design and test a single site-anchored, multiplexed and expandable PCR assay for common types of cutaneous HPVs. STUDY DESIGN: Common forward and unique reverse primers were selected from the E2 open reading frames of five cutaneous HPV genotypes. These were analyzed for sensitivity and selectivity using pHPV plasmids and several control DNAs in an optimized multiplexed assay. This standardized assay was used to analyze human verruca plantaris tissue for genome type and to evaluate the effect of a commonly used treatment protocol. RESULTS: A sensitive, multiplexed PCR assay for human cutaneous HPV genotypes 1a, 2a and 4 was developed. Specific-unique primers and a consensus anchor primer were selected within the HPV E2 region to produce amplicons varying by greater than 100bp. In analytical sensitivity studies, fewer than 100 genome copies of HPV1a and 2a were detected, and fewer than 1000 copies of HPV4 were detected. The multiplexed assay did not amplify regions of human placenta, calf thymus, CaSki or SiHa DNA and E. coli, pBR322 or non-HPV virus DNAs. In combination with a forensic DNA extraction procedure, the multiplexed HPV assay detected and identified HPV types in 23 of 51 (45%) deep plantaris verrucae. Two patients were found with two different genotypes in single deep plantaris verruca. Detection of the HPV genome was followed as a function of tissue ablation and Mediplast treatment in one patient. In healing tissue, the genome content was reduced but had not totally disappeared. CONCLUSIONS: The multiplexed HPV assay can be used to determine genotype prevalence that may correlate with treatment efficacy.

Multicentric endobronchial smooth muscle tumors associated with the Epstein-Barr virus in an adult patient with the acquired immunodeficiency syndrome: a case report.

BACKGROUND: The incidence of benign and malignant smooth-muscle tumors (leiomyomas and leiomyosarcomas) is increased in children with the acquired immunodeficiency syndrome (AIDS). Epstein-Barr virus (EBV) infection has been implicated in the pathogenesis of these tumors. Smooth muscle tumors in adults with AIDS are extremely rare, with only six cases involving extrapulmonary sites reported in the literature. METHODS: Multifocal smooth walled endobronchial tumors were removed from a 35-year-old man with AIDS using rigid bronchoscopic laser resection. The tumor tissues were processed for routine histology, immunohistochemical stainings, and EBV in situ hybridization using an EBV-encoded RNA- 1 RNA oligonucleotide probe. RESULTS: Histologic features and immunohistochemical profiles were characteristic of smooth muscle tumors. EBV gene expression was detected in > 90% of tumor cell nuclei. Although overt histopathologic evidence of malignancy was lacking, some of the histopathologic findings, along with multifocality of the tumors and the rapid appearance of new tumors, suggested an unfavorable prognosis in this case. CONCLUSIONS: To the authors' knowledge, this is the first reported case of multicentric smooth muscle tumors involving the bronchi and lungs of an adult patient with AIDS. Diffuse EBV gene expression in the tumor tissue supports the hypothesis that EBV infection contributes to the pathogenesis of tumors of smooth muscle origin in immunocompromised hosts.