Association of genotype III of dengue virus serotype 3 with disease outbreak in Eastern Sudan, 2019.

BACKGROUND: Dengue fever (DF) is an arthropod-borne disease caused by dengue virus (DENV). DENV is a member of the genus Flavivirus in the family Flaviviridae. Recently, DENV has been reported as an important emerging infectious viral pathogen in Sudan. Multiple outbreaks and sporadic cases of DF have been frequently reported in the eastern region of Sudan. The present study was conducted to confirm DENV outbreak in Kassala State, eastern Sudan, 2019, and to provide some information on the molecular characterization of the DENV isolate associated with the disease outbreak. METHODS: A hundred serum samples were collected during the outbreak from residents of Kassala State, Sudan, 2019. ELISA was used to detect DENV non structural protein NS1 (DENV-NS1) in acute phase sera sampled during the disease outbreak. RT-PCR assays were used to amplify a fragment of the capsid/pre-membrane region (CprM) of the viral polyprotein gene. The PCR products of the amplified CprM region of the viral polyprotein gene were purified and partial sequences were generated and used to confirm the specificity of DENV sequences and to identify the virus serotype. Phylogenetic tree was constructed to determine the genotype of DENV associated with the outbreak. RESULTS: Using DENV-NS1 ELISA assay, DENV infection was confirmed in 23% sampled sera. The detection of DENV RNA was made possible using group-specific RT-PCR assay. The virus was serotyped as DENV serotype 3 (DENV-3) using DENV serotype-specific RT-PCR assay. Phylogenetic analysis of the partial CprM sequences of the viral polyprotein gene indicates that the virus belonged to genotype III of DENV-3. CONCLUSION: The scientific data presented in this investigation confirmed that genotype III of DENV-3 was associated with the disease outbreak in eastern Sudan, 2019. The study represents the first report on molecular characterization of DENV-3 in Sudan.

Recent transmission of dengue virus and associated risk Facors among residents of Kassala state, eastern Sudan.

BACKGROUND: Acute arboviral infections are distributed worldwide including Sudan, and dengue fever (DENV) is not an exception. The virus activity has recently been frequently reported in Kassala State, eastern Sudan. However, an appropriate epidemiological study would be necessary to provide accurate and precise estimates of the magnitude of recent DENV transmission in this area of endemicity. METHODS: In the present investigation, a cross sectional study was conducted to advance beyond the current knowledge of the epidemiology of the disease in Kassala State. The prevalence of the disease was estimated and associated risk factors were determined. Sampled sera were collected and screened for recent dengue transmissionas as determined by DENV-IgM enzyme-linked immunosorbent assay (ELISA). The collection of data for risk assessment was supported by a well designed structured questionnaire. RESULTS: The prevalence of recent DENV infection was estimated to be (11.42%). Potential risk factors to DENV seropsitivity include, age (OR = 3.24, CI = 1.81-5.77,p-value = 0.001); low income (OR = 3.75, CI = 1.57-8.93, p-value = 0.027); mosquito control (OR = 4.18, CI = 2.33-7.51, p-value = 0.004); and localities. CONCLUSION: The present study showed a high rate of circulating DENV IgM antibodies among the participants of the study (11.42%), suggesting recent transmission of DENV in Kassala State, eastern Sudan. The frequent occurrence of DENV infections necessitates the need for improved surveillance programs and prevention measures to combat this important arboviral disease in Sudan.

First molecular characterization of Echinococcus granulosus (sensu stricto) genotype 1 among cattle in Sudan.

BACKGROUND: Echinococcus granulosus sensu lato (s.l.) is the causative agent of cystic echinococcosis (CE), which is a cosmopolitan zoonotic parasitic disease infecting humans and a wide range of mammalian species including cattle. Currently, little information is available on the genetic diversity of Echinococcus species among livestock in Sudan. In the present study, fifty (n = 50) hydatid cysts were collected from cattle carcasses (one cyst sample per animal) at Al-kadarou slaughterhouse, Khartoum North, Sudan. DNA was extracted from protoscolices and the germinal layer of each cyst and subsequently amplified by PCR targeting the mitochondrial NADH dehydrogenase subunit 1 (NADH-1) gene. The amplified PCR products were purified and subjected to direct sequencing for subsequent construction of phylogenetic tree and net work analysis. RESULTS: The phylogenetic tree revealed the presence of Echinococcus canadenesis genotype 6 (G6) in 44 cysts (88.0%), Echinococcus ortleppi genotype 5 (G5) in 4 cysts (8.0%) and Echinococcus granulosus sensu stricto (s.s) genotype 1 (G1) in 2 cysts (4.0%). The phylogenetic network analysis revealed genetic variation among the different haplotypes/genotypes. This report has provided, for the first time, an insight of the role of cattle in the transmission of the zoonotic G1 echinococosis. CONCLUSIONS: The results of the study illustrate that Sudanese breeds of cattle may play an important role in the transmission dynamics and the epidemiology of cystic echinococcosis in Sudan. This study reports the first molecular identification of E. granulosus s.s. in cattle in Central Sudan.

Sero-epidemioloical survey on African horse sickness virus among horses in Khartoum State, Central Sudan.

BACKGROUND: African horse sickness virus (AHSV) is an infectious non contagious insect-transmitted double-stranded (ds) RNA orbivirus of the family Reoviridae. AHSV causes an often fatal hemorrhagic infection with high mortality among selected breeds of Arabian horses. This study was conducted to avail some information with regard to the prevalence and associated risk factors of AHSV among ecotype breeds of horses in central Sudan. METHODS: Sera were collected from 320 horses, which were selected randomly from four localities and employed in the study. A competitive enzyme-linked immunosorbent assay (cELISA) was used to screen sampled sera for AHSV-specific immunoglobulin G (Ig G) antibodies. RESULTS: Seropositivity to AHSV Ig G was detected in 275 out of the 320 horse sera, thus accounting for a prevalence rate of 85.9%. Potential risk factors to AHSV infection were reported to be associated with horse breed (OR = 5.0, CI = 0.07-2.104, p-value = 0.039) and activity of the horse (OR = 3.21, CI = 0.72-1.48, p- value = 0.008). CONCLUSIONS: The high prevalence of AHSV in Khartoum State of Central Sudan necessitates the need for continuous surveillance for AHSV infection to prevent a possible disease outbreak in this region of the African continent.

Prevalence of dengue fever virus antibodies and associated risk factors among residents of El-Gadarif state, Sudan.

BACKGROUND: Dengue fever, caused by dengue virus (DENV), has become one of the most important mosquito-borne viral diseases with a steady rise in global incidence, including the Sudan. Sporadic cases and frequent acute febrile illness outbreaks, compatible with Dengue fever, have been reported in El-Gadarif State, Sudan. However, diagnosis was based almost exclusively on clinical signs without confirmatory laboratory investigations. Despite the magnitude of the problem in El-Gadarif State, no information is currently available with regard to the epidemiology of the disease in this State. El-Gadarif State is one of the largest commercial centers in the Sudan. The objective of the present investigation is to estimate the prevalence of DENV antibodies, and determine the potential risk factors associated with seropositivity among residents of El-Gadarif State. METHODS: A cross sectional study was conducted in a total of 701residents randomly selected from all 10 localities in El-Gadarif State. The sera from the 701 residents were tested for the presence of DENV-specific immunoglobulin G (IgG) antibodies using a commercially available Anti-dengue IgG enzyme-linked immunosorbent assay (ELISA). RESULTS: Among the 701 residents, 334 residents (47.6%) were seropositive for DENV. Mosquito control (OR = 2.73, CI = 1.37-5.87, p-value = 0.001); low income (OR = 2.31, CI: 1.71-6.36, p value = 0.032); sleeping out-doors (OR = 3.73, CI = 2.63-6.23, p-value = 0.013), and localities were determined as potential risk factors for contracting DENV infection. CONCLUSIONS: The prevalence rate of DENV antibodies among residents of El-Gadarif State is significantly high (47.6%). Further epidemiologic studies including, distribution of mosquito vectors and implementation of improved surveillance are urgently warranted for better prediction and prevention of a possible DENV outbreak in El-Gadarif State, Sudan.

Crimean Congo hemorrhagic fever among the one-humped camel (Camelus dromedaries) in Central Sudan.

BACKGROUND: Crimean-Congo hemorrhagic fever (CCHF) is a tick-borne viral zoonotic disease caused by Crimean-Congo hemorrhagic fever virus (CCHFV), a member of the genus Nairovirus in the family Bunyaviridae. CCHF is typically asymptomatic in animals but can be highly fatal in humans approaching case fatality rate of approximately 30%. In the present investigation, a cross sectional study was conducted to determine the prevalence of CCHF and to identify the potential risk factors associated with CCHFV seropositivity among the one-humped camel (Camelus dromedaries) in Central Sudan. METHODS: A total of 361 camels selected randomly from six localities were employed in the study. Sera sampled were tested for the presence of CCHFV-specific immunoglobulin G (IgG) antibodies using enzyme-linked immunosorbent assay (ELISA). RESULTS: CCHFV seropositivity was recorded in 77 out of 361 animals accounting for a prevalence rate of 21.3%. Age (OR = 3.6, CI = 1.72-7.79, p-value = 0.026); locality (OR = 5.85, CI = 1.81-18.83, p- value = 0.003), tick number (OR = 4.6, CI = 1.37-9.81, P-value 0.04); tick control (OR = 2.2, CI, 1.11-4.35, P-value = 0.023) and breed (OR = 6.60, CI = 2.38-18.36, P-value = 0.001) were recorded as potential risk factors for contracting CCHF. CONCLUSIONS: The prevalence of CCHF is significantly high among camels in Khartoum State, Sudan. Age, breed, locality and tick control are considered as potential risk factors for contracting CCHF. This study would be expected to reduce the impact on the livelihood of pastoral communities and ultimately avoid disease spread in human.

Development and evaluation of real-time loop-mediated isothermal amplification assay for rapid detection of cystic echinococcosis.

BACKGROUND: Cystic echinococcosis (CE) or hydatidosis, caused by the larval stage of Echinococcus granulosus (EG)-complex, is a neglected parasitic disease of public health importance. The disease is endemic in many African and Mediterranean countries including the Sudan. The objective of the present study was to develop and evaluate a real-time loop-mediated isothermal amplification (LAMP) assay for simple and rapid detection of CE in humans and domestic live stock in Sudan. METHODS: A set of six LAMP primers, designed from the mitochondrial NADH-1 gene of EG cattle strain of genotype 5 (G5), was used as a target for LAMP assay. The assay was performed at a constant temperature (63 °C), with a real-time follow-up using a LightCycler and fluorochrome dye. Following amplification cycles in a simple water bath, LAMP products were observed for color change by naked eye and were visualized under UV light source using agarose gel electrophoresis. RESULTS: The real-time LAMP assay identified a variety of hydatid cysts strains recovered in the Sudan, including Echinococcus canadenses (G6) and Echinococcus ortleppi (G5). Real-time LAMP positive results were detected by the presence of an amplification curve, whereas negative results were indicated by absence of fluorescence detection. Positive LAMP results appeared as a bluish-colored reaction as observed by naked eye, whereas negative LAMP results were observed as purple-colored reaction. The sensitivity studies indicated that the LAMP assay detected as little as a 10 fg of parasite DNA. There was 100 % agreement between results of the LAMP assay and our previously described nested PCR when testing 10-fold serial dilution of DNA extracted from EG-complex hydatid cyst. However, there was no cross-reactivity with other parasites including cysticercus bovis, Fasciola gigantica, and Schistosoma bovis and nucleic acid free samples. CONCLUSION: The developed LAMP assay would be expected to prove highly significant in epidemiological surveys of CE in developing countries or areas of resource-poor settings for both ease of use and cost.

A survey of rift valley fever and associated risk factors among the one-humped camel (Camelus dromedaries) in Sudan.

BACKGROUND: Rift valley fever (RVF) is a mosquito-borne viral disease of domestic livestock and wild ruminants. In camels RVF may cause abortion among pregnant camels, but is most often asymptomatic among other camels. In this study, a seroepidemiological survey was conducted to determine the prevalence of RVFV antibodies and to identify the potential risk factors associated with RVFV seropositivity among the Sudanese one-humped camel (Camelus dromedaries) in Khartoum State, Sudan. A cross sectional study was conducted in Khartoum State, Sudan, in a total of 240 camels selected randomly from four localities. Sera sampled were tested for the presence of RVFV-specific immunoglobulin G (IgG) antibodies using a competitive enzyme-linked immunosorbent assay (c-ELISA). RESULTS: RVFV seropositivity was recorded in 23 out of 240 animals, prevalence rate of 9.6 % among camels in Khartoum State. Age (OR = 8.29, p-value = 0.04) and heavy rainfall (OR = 5.36, p value = 0.01) were recorded as potential risk factors for contracting RVF. CONCLUSIONS: Older age and heavy rainfall were considered as potential risk factors for seropositivity to RVF. Surveillance for RVF among camels and distribution of mosquito vectors should continue to better understand the clinical signs associated with RVFV infection in camels and provide public health authorities an opportunity to anticipate and prepare for a possible RVF outbreak in Khartoum State, Sudan.

Prevalence of bluetongue virus infection and associated risk factors among cattle in North Kordufan State, Western Sudan.

BACKGROUND: Bluetongue virus causes febrile disease in sheep and a fatal hemorrhagic infection in North American White-tailed deer. However, in cattle the disease is typically asymptomatic and no clinical overt disease is associated with bluetongue infection. Bluetongue virus activity has been detected in Khartoum, Sennar and South Darfur states of the Sudan. Currently, no information is available in regard to previous exposure of livestock to Bluetongue virus in North Kordufan State, the largest livestock producing region in the country. The present study was conducted to determine the prevalence of bluetongue antibodies and to identify the potential risk factors associated with the presence of bluetongue antibodies among cattle in North Kordufan State, Sudan. A total of 299 bovine blood samples were collected randomly from six localities in North Kordufan State and were tested by enzyme-linked immunosorbent assay (ELISA) for detection of BTV-specific immunoglobulin G (IgG) antibodies. RESULTS: The serological evidence of Bluetongue virus infection was observed in 58 out of 299 cows, accounting for a 19.4% prevalence rate among cattle in North Kordufan State. Older cattle (>2 years of age) had four times the odds to be infected with BTV compared to young cattle (OR = 4.309, CI = 1.941-9.567, p-value = 0.01). Application of preventive measures, such as spraying or dipping with insecticide protects cattle against Bluetongue infection. Application of vector control measures decreased the odds for bluetongue seropositivity by 7 times (OR = 7.408, CI = 3.111-17.637, p-value = 0.01). CONCLUSIONS: The results of this study indicated that age and application of routine insecticides are influential risk factors for seroprevalence of Bluetongue in cattle. Surveillance of Bluetongue virus should be extended to include other susceptible animals and to study the distribution of the insect vectors in the region to better predict and respond to BTV outbreak in the State of North Kordufan, Sudan.

Prevalence of bluetongue virus antibodies and associated risk factors among cattle in East Darfur State, Western Sudan.

BACKGROUND: Bluetongue virus (BTV) is an insect-transmitted virus, which causes bluetongue disease (BT) in sheep and a fatal hemorrhagic infection in North American white-tailed deer. However, in cattle the disease is typically asymptomatic and no overt clinical signs of disease appear to be associated with BTV infection. Serological evidence and isolation of different BTV serotypes have been reported in Sudan, however, no information is currently available in regard to previous exposure of Sudanese livestock to BTV infection in East Darfur State, Sudan. AIMS: To determine the prevalence of BTV antibodies and to identify the potential risk factors associated with BTV infection among cattle in East Darfur State, Sudan. METHODS: A total of 224 blood samples were collected randomly from five localities in East Darfur State, Sudan. The serum samples were screened for detection of BTV-specific immunoglobulin G (IgG) antibodies using a competitive enzyme-linked immunosorbent assay (c-ELISA). RESULTS: Serological evidence of BTV infection was observed in 150 out of 224 animals accounting for a 67% prevalence rate among cattle in East Darfur State. Older cattle (>2 years of age) were six times more likely to be infected with BTV (OR = 6.62, CI = 2.87-15.26, p-value = 0.01). Regarding animal source (contact with other herds) as a risk factor, it was shown that cattle purchased from market or introduced from other herds were 3 times at higher risk of being infected with BTV (OR = 3.87, CI = 1.07-13.87, p value = 0.03). Exposure of cattle to the insect vector increased the risk of contracting BTV infection by six times compared to non-exposed cattle (OR = 6.44, CI = 1.53-27.08, p value = 0.01). CONCLUSION: The present study indicated that age, animal source and the intensity of the insect vector are influential risk factors for BTV infection in cattle in the Darfur region. Surveillance for BTV infection should be extended to include other susceptible ruminants and to study the distribution of the insect vectors to better predict and respond to a possible BTV outbreak in the State of East Darfur, Sudan.

Rift Valley fever, Sudan, 2007 and 2010.

To elucidate whether Rift Valley fever virus (RVFV) diversity in Sudan resulted from multiple introductions or from acquired changes over time from 1 introduction event, we generated complete genome sequences from RVFV strains detected during the 2007 and 2010 outbreaks. Phylogenetic analyses of small, medium, and large RNA segment sequences indicated several genetic RVFV variants were circulating in Sudan, which all grouped into Kenya-1 or Kenya-2 sublineages from the 2006-2008 eastern Africa epizootic. Bayesian analysis of sequence differences estimated that diversity among the 2007 and 2010 Sudan RVFV variants shared a most recent common ancestor circa 1996. The data suggest multiple introductions of RVFV into Sudan as part of sweeping epizootics from eastern Africa. The sequences indicate recent movement of RVFV and support the need for surveillance to recognize when and where RVFV circulates between epidemics, which can make data from prediction tools easier to interpret and preventive measures easier to direct toward high-risk areas.

A seroepidemiological survey of Crimean Congo hemorrhagic fever among cattle in North Kordufan State, Sudan.

BACKGROUND: Crimean Congo hemorrhagic fever (CCHF), caused by CCHF virus (CCFV), may cause a fatal hemorrhagic illness in humans with mortality rate of approximately 30%. However, in animals the disease is typically asymptomatic and no clinical hemorrhagic infections appears to be associated with CCHFV. Recently, CCHF activity has been detected in western and southern Kordufan region, Sudan. Currently, no information is available in regard to previous exposure of livestock to CCHFV infection in the region. AIMS: In the present study, a seroepidemiological survey was conducted to determine the prevalence of CCHF and to identify the potential risk factors associated with the disease among cattle in North Kordufan State, Sudan. METHODS: In this survey, 299 blood samples were collected randomly from six localities in North Kordufan State and were tested by enzyme-linked immunosorbent assay (ELISA) for detection of CCHFV-specific immunoglobulin G (IgG) antibodies. RESULTS: The result of the study indicated that the prevalence rate of CCHF was relatively high among cattle, where serological evidence of the infection was observed in 21 (7.0%) of 299 animals. Older cattle were eight times more likely to be infected with the virus (OR=8.0824, CI=1.174-66.317, p-value=0.034). Cross breeds were at 37 time higher at risk compared to endogenous breed (OR=37.06, CI=1.455-944, p-value=0.029). Highly tick-infested cattle are 6 times higher at risk for CCHF when compared to tick-free animals (OR=6.532, CI=1.042-10.852, p-value=0.030). CONCLUSION: It is recommended that surveillance of CCHF should be extended to include other ruminant animals and to study the distribution of ticks in the region to better predict and respond to CCHF outbreak in the State of North Kordufan, Sudan.

First report on circulation of Echinococcus ortleppi in the one humped camel (Camelus dromedaries), Sudan.

BACKGROUND: Echinococcus granulosus (EG) complex, the cause of cystic echinococcosis (CE), infects humans and several other animal species worldwide and hence the disease is of public health importance. Ten genetic variants, or genotypes designated as (G1-G10), are distributed worldwide based on genetic diversity. The objective of this study was to provide some sequence data and phylogeny of EG isolates recovered from the Sudanese one-humped camel (Camelus dromedaries). Fifty samples of hydatid cysts were collected from the one- humped camels (Camelus dromedaries) at Taboul slaughter house, central Sudan. DNAs were extracted from protoscolices and/or associated germinal layers of hydatid cysts using a commercial kit. The mitochondrial NADH dehydrogenase subunit 1 (NADH1) gene and the cytochrome C oxidase subunit 1 (cox1) gene were used as targets for polymerase chain reaction (PCR) amplification. The PCR products were purified and partial sequences were generated. Sequences were further examined by sequence analysis and subsequent phylogeny to compare these sequences to those from known strains of EG circulating globally. RESULTS: The identity of the PCR products were confirmed as NADH1 and cox1 nucleotide sequences using the Basic Local Alignment Search Tool (BLAST) of NCBI (National Center for Biotechnology Information, Bethesda, MD). The phylogenetic analysis showed that 98% (n = 49) of the isolates clustered with Echinococcus canadensis genotype 6 (G6), whereas only one isolate (2%) clustered with Echinococcus ortleppi (G5). CONCLUSIONS: This investigation expands on the existing sequence data generated from EG isolates recovered from camel in the Sudan. The circulation of the cattle genotype (G5) in the one-humped camel is reported here for the first time.

A nosocomial transmission of crimean-congo hemorrhagic fever to an attending physician in North Kordufan, Sudan.

BACKGROUND: Crimean-Congo hemorrhagic fever (CCHF), a tick-borne disease caused by Crimean-Congo hemorrhagic fever virus (CCHFV), is a member of the genus Nairovirus in the family Bunyaviridae. Recently, CCHFV has been reported as an important emerging infectious viral pathogen in Sudan. Sporadic cases and multiple CCHF outbreaks, associated with nosocomial chain of transmission, have been reported in the Kordufan region of Sudan. AIMS: To confirm CCHF in an index patient and attending physician in North Kordufan region, Sudan, and to provide some information on virus genetic lineages. METHODS: Antibody captured ELISA, reverse transcription PCR, partial S segment sequences of the virus and subsequent phylogenetic analysis were used to confirm the CCHFV infection and to determine the virus genetic lineages. RESULTS: CCHF was confirmed by monitoring specific IgM antibody and by detection of the viral genome using RT-PCR. Treatment with oral ribavirin, replacement with fluid therapy, blood transfusion and administration of platelets concentrate resulted in rapid improvement of the health condition of the female physician. Phylogenetic analysis of the partial S segment sequences of the 2 CCHFV indicates that both strains are identical and belong to Group III virus lineage, which includes viruses from Africa including, Sudan, Mauritania, South Africa and Nigeria. CONCLUSION: Further epidemiologic studies including, CCHFV complete genome analysis and implementation of improved surveillance are urgently needed to better predict and respond to CCHF outbreaks in the Kordufan region, Sudan.

Prevalence of cystic echinococcosis and associated risk factors among humans in Khartoum State, Central Sudan.

BACKGROUND: Hydatid disease or cystic echinococcosis (CE) is caused by the larval stages of the cestode parasite Echinococcus granulosus. The objectives of this study were to estimate the prevalence of seropositivity and to identify the risk factors associated with the disease among humans in Khartoum State, Central Sudan. METHODS: A cross-sectional study was conducted between November 2017 and April 2018. A total of 305 randomly selected consenting participants from three localities were included in the current investigation using a multistage probability sampling method. An in-house enzyme-linked immunosorbent assay was used to detect immunoglobulin G antibodies to E. granulosus. The χ2 test and logistic regression analysis were used to determine the risk factors associated with CE seropositivity. RESULTS: A seroprevalence of 6.5% (20/305) was recorded among humans in Khartoum State, Central Sudan. Age (odds ratio [OR] 16.61 [confidence interval {CI} 2.21 to 117.92], p=0.006), locality (OR 3.08 [CI 1.42 to 22.54], p=0.011) and contact with dogs (OR 2.34 [CI 0.026 to 0.646], p=0.013) were recorded as potential risk factors for seropositivity to CE in the study area. CONCLUSIONS: The seroprevalence of CE (6.5%) is high among humans in Khartoum State, Central Sudan. Improved surveillance is necessary to optimize control and prevention strategies for CE as an important neglected zoonotic disease among the human population in the study area of Central Sudan.

Nosocomial outbreak of Crimean-Congo hemorrhagic fever, Sudan.

To confirm the presence of Crimean-Congo hemorrhagic fever in Sudan, we tested serum of 8 patients with hemorrhagic fever in a rural hospital in 2008. Reverse transcription-PCR identified Crimean-Congo hemorrhagic fever virus. Its identification as group III lineage indicated links to virus strains from South Africa, Mauritania, and Nigeria.

Rift Valley Fever among febrile patients at New Halfa hospital, eastern Sudan.

BACKGROUND: Since the first isolation of the Rift Valley Fever virus (RVFV) in 1930s, there have been several epizootics outbreaks in the tropic mainly in Africa including Sudan. Recognition of cases and diagnosis of RVF are critical for management and control of the disease. AIMS: To investigate the seroprevalence and risk factors for seropositive to RVFV IgG among febrile patients. METHODS: All febrile patients presented to New Halfa hospital in eastern Sudan during September through November 2007 were investigated to identify the cause of their fever including malaria and RFV. RESULTS: Out of 290 feverish patients presented to the hospital, malaria was diagnosis in 94 individuals. Fevers of unknown origin were diagnosed in 149 patients. Seropositive to RVFV IgG was detected by enzyme-linked immunosorbent assay in 122 (81.8%) of the sera from these 149 patients with fever of unknown origin. While socio-demographic characteristics (age, Job, education and residency) were not associated with seropositive to RVFV IgG, male (OR = 2.8, 95% CI = 1.0-7.6; P = 0.04) were at three times higher risk for seropositive to RVFV IgG. CONCLUSION: There was a high seropositive to RVFV IgG in this setting, more research is needed perhaps using other methods like PCR and IGM.

A simple and rapid method for detection of Trypanosoma evansi in the dromedary camel using a nested polymerase chain reaction.

A nested polymerase chain reaction (nPCR)-based assay, was developed and evaluated for rapid detection of Trypanosoma evansi in experimentally infected mice and naturally infected camels (Camelus dromedarius). Four oligonucleotide primers (TE1, TE2, TE3 and TE4), selected from nuclear repetitive gene of T. evansi, were designed and used for PCR amplifications. The first amplification, using a pair of outer primers TE1 and TE2, produced a 821-bp primary PCR product from T. evansi DNA. The second amplification, using nested (internal) pair of primers TE3 and TE4, produced a 270-bp PCR product. T. evansi DNAs extracted from blood samples of experimentally infected mice and naturally infected Sudanese breed of dromedary camels were detected by this nested PCR-based assay. The nested primers TE3 and TE4 increased the sensitivity of the PCR assay and as little as 10 fg of T. evansi DNA (equivalent to a single copy of the putative gene of the parasite) was amplified and visualized onto ethidium bromide-stained agarose gels. Amplification products were not detected when the PCR-based assay was applied to DNA from other blood parasites including Thieleria annulata, Babesia bigemina or nucleic acid free samples. Application of this nPCR-based assay to clinical samples resulted in direct detection of T. evansi from a variety of tissue samples collected from experimentally infected mice and blood from naturally infected camels. The described nPCR-based assay provides a valuable tool to study the epidemiology of T. evansi infection in camels and other susceptible animal populations.

Serogrouping of United States and some African serotypes of bluetongue virus using RT-PCR.

The diagnostic potential of RT-PCR for detection of bluetongue virus (BTV) ribonucleic acid (RNA) sequence in cell culture and tissue samples from infected ruminants from United States, Sudan, South Africa and Senegal, was evaluated. The non structural protein 1 (NS1) gene of North American BTV serotype 11 was targeted for PCR amplification. The United States BTV serotypes 2, 10, 11, 13 and 17 and the Sudanese BTV serotypes 1, 2, 4 and 16 and BTV serotype 4 from South Africa and BTV serotype 2 from Senegal were studied. RNAs from all BTV field isolates used in this study, propagated in cell cultures, were detected by the described RT-PCR-based assay. The first specific 790bp BTV PCR products were amplified using a pair of outer primers (BTV1 and BTV2). Specificity of the PCR products was confirmed by a nested amplification of a 520bp PCR product using a pair of internal (nested) primers (BTV3 and BTV4). The BTV PCR products were visualized on ethidium bromide-stained agarose gels. Amplification products were not detected when the RT-PCR-based assay was applied to RNAs from closely related orbiviruses including, epizootic hemorrhagic disease virus (EHDV) prototypes serotypes 1, 2, 4; RNA from Sudanese isolate of palyam orbiviruses serogroup and total nucleic acid extracts from uninfected Vero cells. Application of the nested BTV RT-PCR to clinical samples resulted in amplification of BTV RNA from blood and serum samples from goats experimentally infected with BTV4 and from naturally infected sheep, goats, cattle and deer. The results of this study indicated that this RT-PCR assay could be applied for rapid detection of BTV, in cell culture and clinical samples from susceptible ruminants during an outbreak of the disease, in the United States and African.