We report two cases of confirmed Ebola virus disease in pregnant women, who presented at the Médecins Sans Frontières Ebola treatment centre in Guéckédou. Despite the very high risk of death, both pregnant women survived. In both cases the critical decision was made to induce vaginal delivery. We raise a number of considerations regarding the management of Ebola virus-infected pregnant women, including the place of amniocentesis and induced delivery, and whether certain invasive medical acts are justified.We report two cases of confirmed Ebola virus disease (EVD) in pregnant patients who presented and were treated at the Médecins Sans Frontières (MSF) Ebola treatment centre in Guéckedou. We also raise a number of considerations regarding the role of amniocentesis and induced delivery in the management of pregnant women with EVD.

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A novel method for molecular typing of organisms, amplified fragment length polymorphism analysis, was tested for its suitability in epidemiological studies in medical microbiology. Amplified fragment length polymorphism analysis, originally developed for typing crop plants, consists of a simple restriction-ligation reaction and a subsequent PCR amplification. In a single-step reaction, the genomic DNA is digested and the restriction fragments are ligated to specially constructed adapters. PCR amplification of such tagged restriction fragments with primers complementary to the adapters allows the detection of restriction fragment length polymorphisms upon resolution on agarose gels. The method is fast, efficient, and reproducible for typing strains of Legionella pneumophila isolated from both humans and the environment. The accuracy of the method was tested by comparison with standard restriction fragment length polymorphism typing performed with both a ribosomal and a genomic probe.

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This slaughterhouse study in Chad shows higher proportions of Mycobacterium bovis isolates among Mbororo than Arabe zebu cattle. Spoligotyping shows a homogenetic population structure for M. bovis and lack of spacer 30, as were found in neighboring Cameroon and Nigeria. This finding suggests transborder and ongoing transmission between cattle.

The molecular characterizations of the first 40 Mycobacterium tuberculosis isolates from Chad revealed a high proportion of isolates of the Cameroon family (33%), of which one isolate showed a monodrug resistance. In total, 9/33 (27%) isolates were resistant to isoniazid. The implications of these findings are discussed.In Chad, the annual incidence rate of pulmonary tuberculosis was estimated at 60 to 120 of 100,000 in 1990 (12) but increased to 370 of 100,000 in 2000 (19), making Chad a highincidence country. The recommended WHO treatment strategy for patients with open and extrapulmonary tuberculosis is directly observed chemotherapy, which has been adopted in most African countries, including Chad, where its coverage was 25% in 2003 (20). An increase in drug resistance due to noncompliance during treatment is feared; however, the lack of baseline data on drug resistance from these countries makes monitoring difficult.Drug resistance testing, molecular characterization, and fingerprinting of Mycobacterium tuberculosis complex members have become common practices in most mycobacterial laboratories. A variety of molecular genetic typing tools for M. tuberculosis complex isolates have been developed (18), with the most widely used, IS6110 restriction fragment length polymorphism typing, as the gold standard. More recently, spoligotyping (8) has shown advantages over IS6110 typing. It is more cost-effective, easier to perform, and possible to compare results between laboratories.The outcome of the drug resistance tests and molecular characterization by spoligotyping of the first M. tuberculosis isolates from Chad are presented and implications for treatment control discussed.Between March and July 2001 and February and October 2002, a total of 357 sputum and 282 urine samples were collected from tuberculosis patients at the National Reference Hospital (HGRNT) in the Chadian capital, NЈDjaména, and at four rural health centers that were 50 to 200 kilometers away from NЈDjaména. All specimens (sputum and urine) were decontaminated (9) and inoculated onto two Löwenstein-Jensen slants, one containing 0.75% glycerol and the other containing 0.6% sodium pyruvate. In addition, liquid Middlebrook 7H9 medium containing oleic acid-albumin-dextrose-catalase and polymyxin-amphotericin B-nalidixic acid-trimethoprim-azlocillin was used. The inoculated media were incubated at 37°C without CO 2 for 8 weeks. Smears were made from the sediment and were stained by the Ziehl-Neelsen method (9). Growth of mycobacteria was confirmed by smear. Acid-fast-bacillus-positive colonies were subcultured on three Löwenstein-Jensen slants and a Middlebrook 7H10 agar plate. Three biochemical tests (nitrate, niacin, and 68°C catalase) (9) were used to identify M. tuberculosis complex (MTC) from nontuberculous mycobacteria. The Lebek method was used as an additional phenotypic test to distinguish between MTC members (7), and in addition the standard method for molecular identification of MTC isolates, real-time PCR (10), was performed. Genotyping ...

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Stool specimens from children (<4 years old) with diarrhea were collected over a 1-year period in Ticino (southern region of Switzerland). During the same period, environmental samples were collected from surface waters in the proximity of major water treatment plants. From treatment plants, samples were collected from the raw sewage and before the release of the treated water. From rivers, samples were collected before and after receiving the treated waters. A single-step reverse transcription (RT)-PCR amplification of the entire VP7 gene from extracted double-stranded RNA was developed. For the water samples, a further nested PCR was necessary to increase sensitivity. All amplified viral products were sequenced, and the sequence profile was compared to that of the VP7 genes of human and animal rotaviruses from GenBank. Rotavirus strains are characterized by outer capsid proteins G (glycoprotein) and P (protease-cleaved protein). Correct G genotyping of viral sequences from stool and water samples was possible by analyzing only 189 bp at the 5′ end of the VP7 gene. In the Ticino region, the most predominant G genotype among clinical and water samples was G1. Genotypes G2 and G4 were found only among clinical samples. We also detected rotavirus G1-type sequences in feces from a healthy adult. This finding corroborates the hypothesis that healthy adults act as potential reservoirs for the spread of rotavirus in the environment. In our experiments, this RT-PCR-based method for rotavirus genotyping has proven to be a useful tool for epidemiological investigations.