[Recent smallbox epidemics in Ethiopia and Somalia, human monkey pox virus infections and impossibilities of world eradication of the smallpox].
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The materials of work lasting for 11 years and including the study of 3,425 samples collected at different periods after the eradication of smallpox from patients suspected for orthopoxvirus infection indicate high effectiveness of electron microscopy and especially immune electron microscopy as an integral part of diagnostic examination.
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It cannot be disputed that the WHO plan for the eradication of smallpox has had considerable success. There is no endemic smallpox any longer in the world. Nevertheless caution is advised and continued supervision must not be neglected. There are still countries where supervision is not satisfactory, there is uncertainty about animal reservoirs and it should be assumed from the beginning that up to the present no epidemic disease could be completely eradicated. For this reason it is inevitable that supplies of vaccine should be available. A vaccine from heat sensitive strains is offered which is obtained from cell cultures.
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The presented studies were performed at the WHO Collaboration Center for Smallpox in Moscow in the framework of the WHO monkey pox project. The authors recommend improved methods for rapid detection of orthopoxvirus antigen, namely passive haemagglutination (PHA) using dried stable red blood cells and ELISA in order to provide more rapid and efficient laboratory diagnosis under field conditions. Independent serologic diagnosis of monkey pox by ELISA-adsorption (ELISA-A) was proved of value for epidemiological studies and for detection of inapparent infections. The application range of the latter technique and its limitations were also determined.
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Two fusion experiments with NSO myeloma cells yielded 30 hybrid cell lines which continuously and actively secreted monoclonal antibodies (MoAb) to monkey pox virus. Eight lines appeared to produce antibodies to specific antigenic determinants. The isotype and serologic reactivity of MoAb to monkey pox virus was characterized and the karyotype of several hybridomas determined. An enzyme labelled conjugate has been prepared from MoAb selectively reacting with monkey pox virus. This conjugate allowed the identification of monkey pox virus in the materials from patients. It also helped to determine that an orthopoxvirus isolated from wild squirrel in the Republic of Zaire was monkey pox virus.
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Examinations by EIA of laboratory cultures of various orthopoxviruses and specimens from human-monkey pox cases demonstrated the advantages of a monoclonal preparation over the peroxidase polyclonal conjugate prepared from hyperimmune vaccinia antiserum.
A two-step gel precipitation reaction with antiserum to vaccinia virus, based on sequential addition of two test viruses was elaborated for differentiation of closely related orthopoxviruses. The appearance of an additional precipitation line indicated differences in the antigenic structure of the viruses compared. Differences in the antigenic structure of vaccinia, variola and monkey pox viruses as well as the antigenic identity with cow pox virus of poxviruses isolated from carnivora, elephants and okapi were established.
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An epizootic poxvirus infection occurred in a colony of 80 common marmosets (Callithrix jacchus) recently introduced to a laboratory facility. Over an 18-week period, 29 of the monkeys exhibited skin lesions that persisted for 4-6 weeks. Although eight marmosets died during the outbreak, their deaths were not attributed directly to the poxvirus infection. The skin lesions developed over the entire body surface including the soles and palms. Initially characterized as erythematous papules, they quickly changed to elevated coalescing lesions with extensive scab formation. Histopathologically, the lesions revealed moderate to marked acanthosis, and they progressed to full-thickness epidermal necrosis and ulceration. Intracytoplasmic inclusion bodies were observed occasionally within degenerate keratinocytes. These inclusions most probably constituted the intracytoplasmic aggregates of viral particles observed ultrastructurally and confirmed as members of the poxvirus group by negative staining of direct skin scrapings.
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