Real-time PCR assay for the detection of tanapox virus and yaba-like disease virus.
The yatapoxvirus genus contains three members: tanapox virus (TPV), yaba-like disease virus (YLDV) and yaba monkey tumor virus (YMTV), two of which (TPV and YLDV) may infect humans. However, only a very small number of patients have been diagnosed with TPV outside Africa. Given the increased international travel and the similarity of clinical signs during the early stages of a TPV/YLDV infection as compared to diseases caused by agents of potential biological warfare, such as smallpox, monkeypox, tularemia and anthrax, the rapid and reliable recognition of a TPV/YLDV infection is crucial. A real-time PCR assay using TaqManchemistry was developed in order to identify unambiguously TPV/YLDV. Primers and probe targeting a 101bp region of the PstI L fragment of TPV, initial optimisations steps were carried out with YLDV DNA as template. Using probit regression analysis, the lower limit of detection was calculated to be ca. 8 copies per assay. A total of five TPV strains, one YDLV strain and scab-derived DNA from a patient with a TPV infection yielded specific amplification, whereas the DNA of YMTV was not amplified. Various viral and bacterial pathogens (n=29) associated with rash-causing illnesses were not detected using this assay.
Zimmermann
Pia
P
Bundeswehr Institute of Microbiology, Neuherbergstr. 11, D-80937 Munich, Germany.
Thordsen
Ingo
I
Frangoulidis
Dimitrios
D
Meyer
Hermann
H
D017931:DNA Primers; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D011213:Poxviridae Infections / Q000175:diagnosis*; D013698:Templates, Genetic; D014412:Tumor Virus Infections / Q000175:diagnosis*; D014999:Yaba monkey tumor virus / Q000235:genetics / Q000302:isolation & purification*; D018158:Yatapoxvirus / Q000235:genetics / Q000302:isolation & purification*
D017931:DNA Primers