Accession Number:

ADA428570

Title:

Monkeypox detection in rodents using real-time 3'minor groove binder Taqman assays on the Roche LightCycler, Laboratory Investigation 84:1200 - 1208

Descriptive Note:

Corporate Author:

ARMY MEDICAL RESEARCH INST OF INFECTIOUS DISEASES FORT DETRICK MD

Report Date:

2004-06-21

Pagination or Media Count:

11.0

Abstract:

During the summer of 2003, an outbreak of human monkeypox occurred in the Midwest region of the United States. In all, 52 rodents suspected of being infected with monkeypox virus were collected from an exotic pet dealer and from private homes. The rodents were euthanized and submitted for testing to the United States Army Medical Research Institute of Infectious Diseases by the Galesburg Animal Disease Laboratory, Illinois Department of Agriculture. The rodent tissue samples were appropriately processed and then tested by using an integrated approach involving real-time polymerase chain reaction PCR assays, an antigen-detection immunoassay, and virus culture. We designed and extensively tested two specific real-time PCR assays for rapidly detecting monkeypox virus DNA using the Vaccinia virus F3L and N3R genes as targets. The assays were validated against panels of orthopox viral and miscellaneous bacterial DNAs. A pan-orthopox electrochemiluminescence ECL assay was used to further confirm the presence of Orthopoxvirus infection of the rodents. Seven of 12 58 animals seven of 52 15 of all animals tested positive in both monkeypox-specific PCR assays and two additional pan-orthopox PCR assays in at least one tissue. The ECL results showed varying degrees of agreement with PCR. One hamster and three gerbils were positive by both PCR and ECL for all tissues tested. In addition, we attempted to verify the presence of monkeypox virus by culture on multiple cell lines, by immunohistology, and by electron microscopy, with negative results. Sequencing the PCR products from the samples indicated 100 identity with monkeypox virus strain Zaire-96-I-16 a human isolate from the Congo. These real-time PCR and ECL assays represent a significant addition to the battery of tests for the detection of various orthopoxviruses.

Subject Categories:

  • Medicine and Medical Research
  • Microbiology

Distribution Statement:

APPROVED FOR PUBLIC RELEASE