Identification and typing of molluscum contagiosum virus in clinical specimens by polymerase chain reaction

Abstract

A polymerase chain reaction (PCR) which enables the detection of molluscum contagiosum virus (MCV) genomes in either fresh or formalin-fixed clinical specimens is described. The primers used were designed to amplify a 167 bp region of the 3.8 kbp HindIII fragment K of the MCV 1 genome. The ability of this PCR to detect three common MCV types (1, 1v and 2) in clinical specimens was confirmed using frozen extracts from 75 molluscum lesions, and digests of single sections of 11 formalin-fixed, paraffin-embedded lesions; all of which had been previously typed by Southern hybridisation. In addition, 2 specimens previously negative by hybridisation were shown to be positive for MCV DNA by PCR. Confirmation of the identity of the PCR products and distinction between the two major MCV types (MCV 1/1v versus MCV 2) was achieved by comparison of the results of cleavage with the restriction endonucleases Hhal and Sacl. Sequencing of the PCR products revealed complete homology between MCV 1 and 1v, but minor nucleotide variations between MCV 1/1v and MCV 2 were identified. As well as providing a highly sensitive means of diagnosis, the technique may also prove useful for investigations into the pathogenesis, epidemiology and natural history of molluscum contagiosum infection.

Identification and genotyping of molluscum contagiosum virus from genital swab samples by real-time PCR and Pyrosequencing

“…The use of PCR to identify MCV infection (Nunez et al, 1996;Thompson, 1997) and the use of restriction mapping or hybridization to characterize the subtypes (Darai et al, 1986;Porter et al, 1987;Thompson, 1997) has been previously reported. Although these are functional assays, they are not without their limitations.…”

Molluscum contagiosum simulating a sebaceous cyst: a rare presentation

“…Though this virus cannot be cultured, other diagnostic modalities include electron microscopy, immunohistochemistry using polyclonal antibody and in-situ hybridization for in-situ DNA. 4,8,9 MC presenting as a cutaneous cystic lesion simulating a SC is an extremely rare entity and literature review showed only one such case being reported, to the best of our knowledge. Al-Hilo et al carried out a study on different clinical presentations of MC in Iraqi patients, which demonstrated 28 out of 330 patients (8.48%) with an atypical presentation.…”

Fast and reliable diagnostic methods for the detection of human poxvirus infections

“…Similarly, PCR assays were established that amplified OPV with species-specific amplicon lengths [70]. Some notable conventional PCR assays have been published for other human poxviruses [43,[71][72][73][74][75]]. …”