An 80-Year-Old Man With Respiratory Insufficiency After Intravesical Mycobacterium bovis BCG Immunotherapy.

CASE PRESENTATION: An 80-year-old man came to the ED with fever, hematuria, and overall discomfort for 1 week. His medical history included a superficial urothelial carcinoma of the bladder for which he was adjunctively treated with intravesical Mycobacterium bovis BCG (bacillus Calmette-Guérin) immunotherapy for several months. The patient was admitted to the hospital and was initially treated with cephalosporins for a suspected complicated urinary tract infection, but his symptoms did not improve. Ten days after the initial admission, the patient developed hypoxemic respiratory failure during an episode of fever and cold chills and was admitted to the ICU.

Surveillance technology for HIV-1 subtype C in Ethiopia: an env-based NASBA molecular beacon assay to discriminate between subcluster C and C'.

Forty-nine samples with known C2V3 sequences were used for the evaluation of an env-based molecular beacon assay to distinguish between the two genetic subclusters C and C' which characterize the HIV-1 epidemic in Ethiopia. Two subcluster C and two subcluster C' beacons targeting two different loci in the C2V3 region were developed. Using a three beacon-based (2C and 1C'=C prime), isothermal amplification assay, concordance with DNA sequencing was achieved for 43 (87.8%) samples. Sensitivity was 81.8% and specificity 97.4% for subcluster C beacons. For the subcluster C' beacon, a sensitivity of 97% and a specificity of 87.5% was achieved. Five samples were ambiguous by sequencing of which two samples were subcluster C' by the beacon assay and one subcluster C. Two of the samples remained ambiguous with different beacon-pair combinations as well. From samples with a clear C or C' phylogeny by sequencing, three were undetected by the first-line beacon genotyping assay. Genotype ambiguity was resolved in the three samples using beacon pair combinations restricted to each targeted locus. The beacons were evaluated further in a panel including all HIV-1 subtypes. Four of five subtype C isolates were identified correctly, and no cross-reactivity was observed with other subtypes.

Development of a nucleic acid sequence-based amplification assay that uses gag-based molecular beacons to distinguish between human immunodeficiency virus type 1 subtype C and C' infections in Ethiopia.

A gag-based molecular beacon assay utilizing real-time nucleic acid sequence-based amplification technology has been developed to differentiate between the two genetic subclusters of human immunodeficiency virus type 1 (HIV-1) subtype C (C and C') circulating in Ethiopia. Of 41 samples, 36 could be classified as C or C' by sequencing of the gag gene. All 36 isolates were correctly identified by the gag beacon test. Three isolates with genomes that were recombinant in gag were unambiguously typed as belonging to the C' subcluster. Further analysis revealed that these contained the most sequence homology with a reference subcluster C' sequence in the target region of the beacon and hence were correct for the analyzed region. For one sample, sequencing and gag molecular beacon results did not match, while another isolate could not be detected at all by the beacon assay. Overall, high levels of sensitivity and specificity were achieved for both beacons (90.5% sensitivity and 100% specificity for the C beacon and 100% sensitivity and 95.2% specificity for the C' beacon). The availability of a diagnostic test which can quickly and reliably discriminate between C and C' HIV-1 infections in Ethiopia is an important first step toward studying their respective biological characteristics. As the assay is specific to the Ethiopian HIV-1 subtype C epidemic, it will contribute to characterizing the circulating viruses in this population, thereby generating the information necessary for the development of a potential efficacious HIV-1 vaccine appropriate for the Ethiopian context.

Overdiagnosis of amoebiasis in the absence of Entamoeba histolytica among patients presenting with diarrhoea in Wonji and Akaki, Ethiopia.

To confirm the high reported incidence of intestinal amoebiasis among study participants at 2 cohort sites in Ethiopia where an HIV/AIDS study is taking place, stool samples of 232 patients with complaints of diarrhoea were examined for the presence of Entamoeba histolytica and E. dispar DNA between April and December 2001. By microscopy, 91 (39%) of the study participants were reported to harbour Entamoeba trophozoites and/or four-nucleated cysts. Using specific E. histolytica and E. dispar DNA amplification and detection, none of the study participants were found to be infected with E. histolytica and only 21 (9%) with E. dispar. The consequences of the overdiagnosis of E. histolytica are briefly discussed.

Evaluation of rapid assays for screening and confirming HIV-1 infection in Ethiopia.

To evaluate a simple and rapid testing strategy to diagnose HIV infection in Ethiopia, we subjected a panel of 688 sera with known HIV serologic status (confirmed by ELISA/WB or double ELISA) to 3 rapid assays: Determine HIV-1/2, Capillus HIV-1/2 and Serocard HIV. Samples were obtained from participants in a cohort study on HIV-infection (72%), from tuberculosis patients (18%) and from participants in surveillance studies among police recruits and commercial sex workers (10%). The panel consisted of 249 HIV-1 positive samples, of which 68 were HIV-1 subtype C and 1 HIV-1 subtype A, and 439 HIV-1 negative samples. Determine and Capillus were 100% sensitive and 99.8% specific, Serocard was 100% sensitive and specific. On retrospective evaluation, both parallel (samples tested simultaneously by two rapid assays) and serial (samples tested by two consecutive rapid assays) testing algorithms were 100% sensitive and specific when compared to ELISA/WB or double ELISA testing strategy. In conclusion rapid assays have high sensitivity and specificity. HIV serodiagnosis based on rapid assays may therefore be a valuable alternative in voluntary counselling and testing centres and in facilities where sophisticated laboratories are not available.