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Isolates were genetically related with a small genetic diversity and were positive for ctxA, tcpA El Tor virulence genes. Conclusion The V. Most of the isolates were susceptible to the antibiotic regimen currently used in Tanzania.

The high resistance rate detected for the other common antibiotics calls for continuous antimicrobial susceptibility testing during outbreaks. This is an open access article distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Data Availability: All the relevant data are within the paper and its Supporting information files.

Funding: The author s received no specific funding for this work. Competing interests: The authors have declared that no competing interests exist. Introduction Cholera, a disease caused by V. Out of serogroups of V. About 1. Pathogenic V.

Classical biotype was responsible for most of the epidemic diseases in the past century and was replaced by El Tor biotype, responsible for cholera epidemic in endemic areas. The two biotypes of V. Variants of V. Since when the first case of cholera was reported in Tanzania [ 9 ], the country has experienced numerous cholera outbreaks resulting in many cases and deaths.

Throughout this time, the country continued reporting cholera cases. In approximately 28, cholera cases and deaths with a case fatality rate of 1. Hence, this study aimed to determine the genetic relatedness, virulence factors and antimicrobial susceptibility patterns of V. Methods Study design and setting A cross-sectional study involving eleven regions that experienced cholera outbreaks from January to December was carried out. The study included 99 V. A laboratory identification number was assigned to each specimen to maintain patient confidentiality.

Isolation and identification of V. Forty-nine preserved V. Diagnosis was done in the respective endemic area by experienced personals. Patients with typical symptoms but repeated negative smear or culture results were defined as unconfirmed VL VLS. Sera of these patients reacted with the 14 and 16 kDa antigens, common in all Leishmania species [ 22 ]. Serum samples have been collected on verbal consent of patients. Both proteins, rKLO8 and rK39 antigens, were expressed as 6 x His-tagged fusion proteins and contain the same number of repeats 6 copy.

Individual sera were tested in duplicates and means were taken. Samples with inconsistent results were repeated. Known positive and negative sera were included as controls. A cut off value 0. The antigen was reconstituted by adding 2. Sera were tested at two-fold serial dilutions, ranging from 1: 50 to The DAT titre is indicated as the highest dilution at which agglutination was still visible.

The test device was pre-coated with L. This reaction is indicated by development of a specific colour. Tests were considered positive when purple control and test bands appeared. A single band in the control area indicates a negative result. The test is based on detection of anti-Leishmania antibodies using rKE16 antigen of L.

The antigen is immobilised onto a permeable Nitrocellulose membrane and used to capture anti-Leishmania antibodies present in patients samples. Tests were performed and interpreted as described by the manufacturer. Tests were read within 10 minutes. Sera were considered positive if two magenta red dots, one for Control and the other for Test, appeared.

One dot in the Control area indicates a negative result. The test was considered invalid if no dot appeared in the Control area. These tests rely on antigens of different Leishmania parasites S2 Table. A panel of sera obtained from Sudan, India and France were tested. In addition, 24 sera from symptomatic cases with unconfirmed diagnosis of VL VLS and 25 from asymptomatic cases from the same endemic areas were included.

ELISA tests based on rKLO8 or rK39 were compared quantitatively to assess the performance for detection of anti-Leishmania- specific antibodies in the different regions. Cut off values were calculated for each protein by using 30 sera of healthy non-infected individuals. Sensitivity, specificity, positive predictive value PPV and negative predictive value NPV of each test were calculated and compared in the different endemic regions.

Protein concentrations of 5 ng and serum dilutions of were used as previously determined [ 8 ]. The results Fig. Out of 26 VL sera from India, 25

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