Antigen sandwich ELISA predicts RT-PCR detection of dengue virus genome in infected culture fluids of Aedes albopictus C6/36 cells.

Antigen detection by sandwich ELISA was evaluated to predict RT-PCR detection of dengue viral genome in infected culture fluid of Aedes albopictus clone C6/36 cells. Serum specimens collected from dengue patients within 5 days from onset of fever in 2 hospitals in Metro Manila, Philippines, were inoculated into C6/36 cells, and incubated at 28 degrees C. A total of 282 infected culture fluid specimens were harvested and examined by sandwich ELISA and RT-PCR to detect dengue viral antigen and genome, respectively. In the sandwich ELISA, the P/N ratio was calculated by dividing optical density (OD) of a given test specimen by the OD of the standard negative specimen. Samples with a P/N ratio > or = 4.001 were positive for viral genome detection by RT-PCR. The sensitivity and specificity of antigen sandwich ELISA with RT-PCR as the standard, were 90.4% and 100%, respectively. Although antigen sandwich ELISA is less sensitive than RT-PCR, its usefulness lies in its capability to screen a large number of samples at a minimum cost, especially during an outbreak. Samples that meet a set cutoff value can undergo confirmation by RT-PCR for further epidemiological studies.

Abundance and distribution of Xenopsylla cheopis on small mammals collected in West Java, Indonesia during rodent-borne disease surveys.

During February 2004 and September 2005, Xenopsylla cheopis were collected from small mammal hosts during rodent-bone disease surveys in Jakarta and Bandung, Indonesia. During 4 trap nights in Jakarta, 4 rodent species (Rattus exulans, Rattus norvegicus, Rattus tanezumi and Mus musculus) and one shrew species (Suncus murinus) were collected. Rattus tanezumi had the highest X. cheopis load (128 X. cheopis from 84 R. tanezumi) but R. norvegicus had the highest flea index, 1.8. In Bandung, over 6 trap nights 3 rodent species were collected (R. norvegicus, R. tanezumi and M. musculus) and the shrew, S. murinus, were collected. Rattus norvegicus had the highest number of X. cheopis collected (407 X. cheopis from 181 R. norvegicus) but R. tanezumi had the highest flea index, 3.5. During both surveys, X. cheopis was the species of flea collected.

Bartonella species in rodents and shrews in the greater Jakarta area.

In February 2004, we captured 221 rodents and shrews in the Greater Jakarta area as part of a study to determine the prevalence of rodent-associated vector-borne infections. Microscopic examination of blood smears revealed 6% (13/218) to be positive for Bartonella spp. The corresponding DNA samples, either from blood blots or frozen spleen pieces and from fleas collected on these animals, were tested for evidence of Bartonella infection by PCR, targeting the portions: 378bp and 930bp of the citrate synthase gene (g/tA). The sequences from our sample clusters with a Peruvian entity, B. phoceensis, B. rattimassiliensis and B. elizabethae, the latter species has been associated with endocarditis and neuroretinitis in humans. As previous analyses have shown, there appears to be little geographic or host consistency with phylogenetic placement. The public health significance of these findings remains to be determined.