Sudanese Strains Research Articles

Detection of X-Chromosome Inversions in Anopheles Pharoensis Collected from Egypt and Sudan by PCR Assay

Globally, the WHO estimated 241 million malaria cases with 627000 deaths in Africa in 2020. Studying inversions in the X-chromosomes could detect behavioral changes and efficiency of transmission of Malaria in Anopheles phronesis species. These changes may lead to shifting from becoming a major vector of Malaria after the inability to transmit Malaria. The knowledge of the malaria transmission pattern of Anopheles pharoensis in Sudan needs to be improved. We aimed to detect the presence of any inversions or mutations in Anopheles pharoensis species collected from Egypt and Sudan and to undergo a Polymerase Chain Reaction (PCR) technique to detect mutations or X-chromosome inversions in the Sudanese and Egyptian strains of Anopheles pharoensis also to compare between the X-chromosome of the two species. It is an experimental, analytical study that aimed to detect inversions in the X –chromosomes of Anopheles phronesis collected from Egypt (Faiyoum Governate) and Sudan (Khartoum, Gezira, and Sennar states) of Sudanese Mosquito species. From the gel electrophoresis and PCR assay, inversions were detected by the 100Bp PCR products using primers. Only very few revealed bands from 75 Egyptian strains were detected by bands. The study revealed that Anopheles pharoensis species collected from Egypt have X- chromosome inversions. Inversions were absent in the Sudanese strains. The study also reacted to the fact that Anopheles phronesis could be a sibling species or a species complex, but further studies are needed to prove this. From the results and observations of this investigation, it was recommended to undergo further genetic studies on Anopheles species in Sudan and use of PCR technique in the classification.

Ebola and the Ravages of History in Paul Farmer: A Catholic Theological Ethical Response to Global Health Inequity in Africa

In this chapter, I explore with Paul Farmer why Ebola has persisted in Africa and why it continues to kill so many Africans as we see in the current outbreak in Uganda (September 2022) with the Sudanese strain. Farmer’s appeal to social medicine and social context in understanding the remote causes of the cause of Ebola offers an important corrective to the disease control and treatment paradigm approach in global health amidst the devastating impact of health inequity. I conclude my chapter by using Farmer’s analysis of the causes of global health inequity in Africa to propose how theological ethics can serve the mission of churches and African communities in designing healthcare systems for health improvement and health protection in solidarity with the poor and the most vulnerable. I will propose some effective strategies for global health partnerships in Africa against the current preoccupation with disease control and prevention, medical mission, and crisis intervention from outside Africa.

Molecular characterization of circulating Foot and mouth disease virus (FMDV) serotype O topotype EA-3 and serotype A (African topotype) genotype IV in Egypt, 2016

In January–April 2016, cattle and buffalo farm owners and veterinarians reported clinical signs suggestive of foot and mouth disease virus (FMDV) outbreaks among non-vaccinated cattle and buffalo herds in Egypt. The clinical disease observed was either mild (small oral lesions and speedy recovery) or severe (extensive oral lesions and/or mortalities), and the form of the disease (either mild or severe) segregated by farm. This study aimed to confirm the presence of FMDV and to characterize the circulating strains associated with the outbreaks. Vesicular epithelia were collected from 41 animals representing 15 affected cattle and buffalo farms in five governorates (Behira, Cairo, Daqahlia, Giza and Ismailia), and tested by real time (rt) RT-PCR. Consequently, 92% (38/41) of examined samples were positive. Furthermore, the VP1 coding region of 60% (23/38) of positive specimens were amplified by RT-PCR and sequenced. The phylogenetic analysis identified two distinct strains characterized as serotype O topotype EA-3 and serotype A (African topotype) of genotype IV in the severe and mild disease forms, respectively. The newly identified strains clustered in distinct clades in the phylogenetic trees, indicating the likelihood of new incursions into Egypt. Those strains were most closely related to previously described Sudanese strains.

ASingleLineageofHepatitisEVirusCausesBoth OutbreaksandSporadicHepatitisinSudan.

Few studies have reported sporadic hepatitis E virus (HEV) infections during non-outbreak periods in Africa. In this study, the prevalence of HEV infection in Sudan was investigated in 432 patients with acute hepatitis from 12 localities in North Kordofan, and from 152 patients involved in smaller outbreaks of hepatitis in the neighbouring Darfur. HEV infection was diagnosed in 147 (25%) patients: 98 from Kordofan and 49 from Darfur. The mortality was 10%; six of the patients who died from the infection were pregnant women. HEV RNA was detected by quantitative real-time polymerase chain reaction (RT-qPCR) in 38 (26%) patients: 22 from Kordofan and 16 from Darfur. Partial open reading frame (ORF) 1 and ORF2 were sequenced from HEV from nine and three patients, respectively. Phylogenetic analysis showed that the Sudanese strains belonged to genotype 1 (HEV1), and confirmed the segregation of African HEV1 strains into one branch divergent from Asian HEV1. It also revealed that the Sudanese strains from this study and from an outbreak in 2004 formed a separate clade with a common ancestor, distinct from strains from the neighbouring Chad and Egypt. This HEV strain has thus spread in a large area of Sudan, where it has caused both sporadic hepatitis E and outbreaks from at least 2004 and onwards. These data demonstrate that hepatitis E is a constant, on-going public health problem in Sudan and that there is a need for hepatitis E surveillance, outbreak preparedness, and general improvements of the sanitation in these remote areas of the country.

Detection and Molecular Characterization of Respiratory Syncytial Virus (RSV) in Children with Respiratory Signs in Khartoum State, Sudan 2011-2012

Background: The present study was to investigate the incidence of the respiratory syncytial virus infection in children and to characterize the RSV circulating in Khartoum state during 2011-12 winter seasons. Methodology: Throat swab specimens collected from 224 children less than 5 years old, with respiratory tract infections admitted at Khartoum Hospitals in winter season (2011- 2012), were screened for RSV using direct immunofluorescence assay (DFA) and reverse transcription- polymerase chain reaction (RT-PCR). Isolation in cell culture followed by nucleotide sequencing and bioinformatics analysis based on the G gene, were done for the RT- PCR positive RSV samples. Results: Out of 224 patients, RSV infections were detected in 136 (60.7%) patients, by using DFA technique, and 44 (19.6%) patients using RT-PCR. 22 strains of RSV were isolated in Hep-2 cell line. The clinical symptoms including Bronchiolitis, Pneumonia, Asthma and Allergy showed significantly different rates (p<0.05) in having RSV infection, (P-value = 0.017, 0.002, 0.0001, 0.0001) respectively. Bioinformatics analysis of nucleotide sequences of 7 cell culture isolated RSV strains revealed that all analyzed RSV belonged to the RSV-A genotype. Phylogenetic tree of RSV-A sequences showed that, all Sudanese strains were grouped with strains from Belgium and Saudi Arabia. Conclusions: This is the first report on molecular characterization that describes the circulation of RSV genotype in Sudan. DFA and RT-PCR offers rapid methods for detection of RSV in hospitalized children with Respiratory tract infection (RTI).

Emergence of peste des petits ruminants virus lineage IV in Ismailia Province, Egypt

Peste des petits ruminants (PPR) is an acute, highly contagious fatal disease of small ruminants characterized by high fever, ocular and nasal discharge, pneumonia, erosive stomatitis and severe enteritis that ultimately results in high mortalities. Peste des petits ruminants virus (PPRV) is widely distributed and endemic in several African, middle eastern and south Asian countries and it poses a threat to European countries. Egyptian veterinary medical authorities stated that Egypt is free from PPRV and the only measures for disease control are test and slaughter of infected population to maintain the free status. The aim of our investigation was to detect PPRV in Ismailia province as an indicator of the infection status in Egypt and perform molecular characterization of the emerging virus to gain insight into the origin of circulating virus. A total of 40 representative clinical samples, from a single goat case and goat flock in 2010 and sheep flock in 2012, were tested for PPRV by RT-PCR. About 21 (52.5%) samples were positive. The phylogenetic analysis of the detected viruses revealed circulation of PPRV lineage IV. The circulating viruses are closely related to Sudanese and Saudia Arabian strains with nucleotide identity ranged from 99.2% to 99.6%, respectively. Also, it is closely related to Moroccan 2008 viruses with identities ranged from 97.6% to 98%. Epidemiological investigation at the national level is recommended for monitoring PPRV spread and implementing an appropriate control program.

Test of Cross Reaction Between the Two Local Sudanese Strains of P. mulocida (B and E)

The present study was carried out on the two local Sudanese Pasteurella multocida strain B and E which cause Hemorrhagic Septicemia (HS) disease in the Sudan and used for the production of the vaccine against this disease. This study was done to test the cross reaction between the two strains. The study showed that there was a cross reaction between the two strains in the rabbit sera when tested by ELISA. The active cross protection test in rabbits showed that vaccine of strain E protected 50% of the rabbits against challenge with strain B, while the vaccine of B could not protect the rabbits against the challenge with strain E 0%. Each vaccine of B and E could protect the rabbits 100% against the challenge with the same strain (homologously).

Polymorphism in the HASPB Repeat Region of East African Leishmania donovani Strains

Background/ObjectivesVisceral leishmaniasis (VL) caused by Leishmania donovani is a major health problem in Ethiopia. Parasites in disparate regions are transmitted by different vectors, and cluster in distinctive genotypes. Recently isolated strains from VL and HIV-VL co-infected patients in north and south Ethiopia were characterized as part of a longitudinal study on VL transmission.Methodology/Principal FindingsSixty-three L. donovani strains were examined by polymerase chain reaction (PCR) targeting three regions: internal transcribed spacer 1 (ITS1), cysteine protease B (cpb), and HASPB (k26). ITS1- and cpb - PCR identified these strains as L. donovani. Interestingly, the k26 - PCR amplicon size varied depending on the patient's geographic origin. Most strains from northwestern Ethiopia (36/40) produced a 290 bp product with a minority (4/40) giving a 410 bp amplicon. All of the latter strains were isolated from patients with HIV-VL co-infections, while the former group contained both VL and HIV-VL co-infected patients. Almost all the strains (20/23) from southwestern Ethiopia produced a 450 bp amplicon with smaller products (290 or 360 bp) only observed for three strains. Sudanese strains produced amplicons identical (290 bp) to those found in northwestern Ethiopia; while Kenyan strains gave larger PCR products (500 and 650 bp). High-resolution melt (HRM) analysis distinguished the different PCR products. Sequence analysis showed that the k26 repeat region in L. donovani is comprised of polymorphic 13 and 14 amino acid motifs. The 13 amino acid peptide motifs, prevalent in L. donovani, are rare in L. infantum. The number and order of the repeats in L. donovani varies between geographic regions.Conclusions/SignificanceHASPB repeat region (k26) shows considerable polymorphism among L. donovani strains from different regions in East Africa. This should be taken into account when designing diagnostic assays and vaccines based on this antigen.

Molecular characterization of rabies virus isolated from dogs in Tunisia: Evidence of two phylogenetic variants

In an attempt to explain temporal and geographical rabies incidence fluctuations in Tunisia, a molecular epidemiological study of rabies virus (RV) was carried out. A panel of RV isolates from dogs, collected between 1992 and 2003, from different regions in Tunisia have been analysed by direct sequencing of PCR-amplified products coding for the nucleoprotein gene. New sequences have been compared to prototype sequences of Lyssavirus species and nine lineages of species 1. All Tunisian isolates belonged to species rabies virus and segregated into two rabies lineages geographically distinct: NCS lineage characterizing Northeast, Central and Northern areas of the country and NW lineage more restricted to the North-Western regions. Phylogenetic analyses showed that Tunisian RV clustered most closely to Africa 1a lineage: NCS lineage showed nucleic affiliation with isolates from Algeria and Morocco, whereas, NW lineage shared a strong relationship with Ethiopian and Sudanese strains.

Leishmania donovani complex (Kinetoplastida, Trypanosomatidae): Comparison of deoxyribonucleic acid based techniques for typing of isolates from Ethiopia

In Ethiopia, visceral leishmaniasis (VL) is an increasing public health concern. Recently, a new outbreak of VL claimed the lives of hundreds of Ethiopians. Mapping its distribution and the identification of the causative Leishmania species is important for proper use of resources and for control planning. The choice of appropriate typing technique is the key for determining the infecting species. Here we compared three deoxyribonucleic acid (DNA) based markers. We used, for the first time, cpbE and cpbF (cpbE/F) PCR-RFLP and demonstrated that it clearly differentiates Leishmania donovani from Leishmania infantum. The cpbE/F PCR-RFLP gave identical banding pattern for all L. donovani strains irrespective of their geographic origin. With the K26 (primers) PCR-RFLP, the L. donovani strains gave a banding pattern different from L. infantum and showed variation with geographic origin. The Ethiopian isolates typed as L. donovani by the PCR-RFLP of the cpbE/F (gene) and K26 (primers) showed two types of patterns with the T2/B4 (primers) PCR-RFLP; one group with L. infantum-like and the other L. donovani-like pattern. Phylogenetic analysis using cpbE/F sequences showed variation with geographic origin of strains and the African strains of L. donovani are more distantly related to L. infantum. Moreover, the Ethiopian isolates were seen to be closely related to the Sudanese, Kenyan and Indian strains. Thus, we recommend the use of more than one marker to study the population genetics of L. donovani complex.

Phylogenetic analysis of rabies viruses from Sudan provides evidence of a viral clade with a unique molecular signature

Rabies is endemic in Sudan and remains a continual threat to public health as transmission to humans is principally dog-mediated. Additionally, large-scale losses of livestock occur each year causing economic and social dilemmas. In this study, we analysed a cohort of 143 rabies viruses circulating in Sudan collected from 10 different animal species between 1992 and 2006. Partial nucleoprotein sequence data (400 bp) were obtained and compared to available sequence data of African classical rabies virus (RABV) isolates. The Sudanese sequences formed a discrete cluster within the Africa 1a group, including a small number of sequences that clustered with sequences from Ethiopian RABV. These latter sequences share an Aspartic Acid at position 106 (Asp 106) with all other Africa 1a group members, in contrast to the remaining Sudanese strains, which encode Glutamic Acid at this position (Glu 106). Furthermore, when representatives of other African and European lineages were aligned, Glu 106 is unique to Sudan, which supports the concept of a single distinct virus strain circulating in Sudan. The high sequence identity in all Sudanese isolates studied, demonstrates the presence of a single rabies virus biotype for which the principal reservoir is the domestic dog.

Evaluation of oral therapy on Mansonial Schistosomiasis using single dose of Balanites aegyptiaca fruits and praziquantel

The efficacy of Balanites aegyptiaca fruit mesocarp was compared with praziquantel in mice infected with Sudanese strain of Schistosoma mansoni. Infected mice were given a single dose of 200 mg/kg body weight of B. aegyptiaca fruit mesocarp and 200 mg/kg b.w. of praziquantel after 6 weeks from the onset of the infection. A significant reduction was observed in EPG (egg count per gram of faeces), eggs burden in tissues and recovery of adult worms ( P<0.05) for both the plant and the drug-treated animals.

Leishmania parasites (Kinetoplastida: Trypanosomatidae) reversibly inhibit visceral muscle contractions in hemimetabolous and holometabolous insects

Female sand flies can acquire protozoan parasites in the genus Leishmania when feeding on an infected vertebrate host. The parasites complete a complex growth cycle in the sand fly gut until they are transmitted by bite to another host. Recently, a myoinhibitory peptide was isolated from Leishmania major promastigotes. This peptide caused significant gut distension and reversible, dose-dependent inhibition of spontaneous hindgut contractions in the enzootic sand fly vector, Phlebotomus papatasi. The current study further characterizes myoinhibitory activity in L. major and other kinetoplastid parasites, using the P. papatasi hindgut and other insect organ preparations. Myoinhibitory activity was greatest in cultured promastigotes and in culture medium in late log-phase and early stationary-phase, coinciding with development of infective Leishmania morphotypes in the sand fly midgut. L. major promastigote lysates inhibited spontaneous contractions of visceral muscle preparations from hemimetabolous (Blattaria and Hemiptera) and holometabolous (Diptera) insects. Inhibition of visceral muscle contractions in three insect orders indicates a conserved mode of action. Myoinhibitory activity was detected also in Leishmania braziliensis braziliensis, a Sudanese strain of Leishmania donovani, and the kinetoplastid parasite Leptomonas seymouri. Protozoan-induced myoinhibition mimics the effect of insect myotropins. Inhibiting host gut contractions protects Leishmania parasites from being excreted after blood meal and peritrophic matrix digestion, allowing development and transmission of infective forms.

A neglected disease of humans: A new focus of visceral leishmaniasis in Bakool,Somalia

Visceral leishmaniasis (VL) was observed in children in Bakool region, Somalia, an area where VL hasnot been reported before. We describe the extent of the problem in this war- and famine-stricken area. A retrospective analysis was done of all cases admitted to a VL treatment centre between July 2000 and August 2001. Patients with longstanding fever, splenomegaly and a positive direct agglutination test (DAT; titre > 1:3200) were treated as suspected VL cases. A rapid epidemiological and entomological assessment was performed in the area. Species identification was attempted from blood samples by polymerase chain reaction-restriction fragment length polymorphism analysis of cysteine proteinase B genes. In 1 year, 230 serologically-positive cases were diagnosed as VL, and response to therapy was good in 91.6% of the 225 treated with sodium stibogluconate. Parasitological confirmation was attempted and obtained in 2 cases. Parasites were found to be most similar to Sudanese and Ethiopian reference strains of the Leishmania donovani complex. In a serological survey of 161 healthy displaced persons, 15% were positive by the leishmanin skin test and 3 (2%) were positive by the DAT. The sandfly captures showed Phlebotomus martini and P. vansomerenae. VL seems to be a longstanding and serious health problem in Bakool region. Food insecurity might have contributed to the emergence and detection of VL in this area.

Strain typing in Leishmania donovani by using sequence-confirmed amplified region analysis

To differentiate strains of Leishmania donovani, allelic markers at the DNA level were developed by sequence-confirmed amplified region analysis (SCAR). Homologous fragments from different strains of L. donovani were amplified by PCR using random primers and subsequently screened for single-strand conformation polymorphisms. Direct sequencing revealed 55 sequence polymorphisms in eight co-dominant DNA markers; 38 of them were single point mutations. Heterozygosity was evident for 69% and fixed heterozygosity for 25% of all polymorphisms. At most polymorphic sites one of the segregation genotypes was missing. Nineteen unique multilocus genotypes were identified among 29 strains of L. donovani. One genotype was represented by eight Sudanese strains; also two strains from Sudan as well as two strains from Kenya, respectively, shared identical genotypes. All other strains had individual multilocus genotypes. Calculation of genetic distances showed a correlation between multilocus genotypes and the geographical origin of these strains. African strains were found in one well-supported cluster with Kenyan and Sudanese strains clearly separated. SCAR markers seem to represent a random sample of neutral genetic variation present in natural populations. They are co-dominant because they can detect all possible allele combinations in a diploid organism and may, therefore, be very useful for population genetic analysis in Leishmania.

Characterization of attenuated Theileria annulata vaccines from Spain and the Sudan.

Theileriosis caused by Theileria annulata can be effectively prevented by vaccination with attenuated, cultured schizonts. Although these attenuated vaccines have been applied for a long time, not much is known about the fate of the vaccine strain in the field. Here, two experimental Spanish vaccine strains originating in Cádiz and Cáceres, and one Sudanese strain are studied to address the development of a carrier status and the infectivity for Hyalomma ticks. Moreover, the heterogeneity of the merozoite surface protein, Tams1, was analyzed in search for an attenuation marker. Using the sensitive reverse line blot (RLB) hybridization, the development of a low level carrier status was demonstrated in the Cáceres and Sudanese line vaccinated calves. Although no signal was detected in the Cádiz line vaccinated calves, seroconversion against the schizont stage was observed, as it was in all other calves. The experimental transmission of T. annulata by Hyalomma ticks to naïve calves was unsuccessful for all cell line inoculated calves. Tams1 heterogeneity indicated a clonal selection of parasites during the process of attenuation, but the Tams1 sequence itself has no connection with the attenuation status. In conclusion, a carrier status develops in attenuated schizont culture vaccinated calves, but is not infective for Hyalomma ticks. Based on these data, the risk for spread of the vaccine strains in the field may be very low.

Characterization of the Glucosyltransferases That Assemble the Side Chains of the Indian Leishmania donovani Lipophosphoglycan

The life cycle of Leishmania parasites within its sand fly vector involves the development of extracellular promastigotes from a noninfective, “procyclic” stage into an infective, “metacyclic” stage that is adapted for transmission in the fly and survival in the mammalian host. Lipophosphoglycan (LPG), the predominant surface glycoconjugate in both procyclic and metacyclic stages, is a critical virulence determinant. LPG is a multidomain molecule; the structural polymorphisms among species lie in branching from the backbone 6Galβ1,4Man(α1)-PO4 repeat units and in the composition of the small oligosaccharide caps. We have recently demonstrated that the LPG from an Indian isolate of Leishmania donovani differs from a Sudanese strain by possessing one or two side chain β(1,3)-linked glucose residues. We now have characterized the glucosyltransferase activities responsible for glucosylating the LPG. When incubated with UDP-[3H]glucose and Mn2+, microsomal membranes from the Indian isolate transferred [3H]glucose to the repeat units of the exogenous acceptor Sudanese L. donovani LPG, which does not contain any side chain branching. Glucose addition was maximal at 28°C, the optimal growth temperature of procyclic L. donovani. Consistent with the lack of side chain branching in its LPG, Sudanese L. donovani showed minimal glucosyltransferase activity. Indian metacyclic promastigotes, in contrast to procyclic promastigotes, express no glucose side chains off the repeat units. Therefore, we compared the relative activity of the glucosyltransferases in microsomes from procyclic and metacyclic promastigotes and observed approximately 80% less activity in the latter. These results provide evidence that the glucose side chain addition to LPG is developmentally regulated during the parasite's life cycle and that the glucosyltransferases of L. donovani are strain specific.

Rhizobia Isolated from Root Nodules of Tropical Leguminous Trees Characterized Using DNA-DNA dot-blot Hybridisation and rep-PCR Genomic Fingerprinting

Fifty-one fast growing rhizobial strains isolated from root nodules of Acacia Senegal and Prosopis chilensis in Sudan and Kenya were divided into DNA homology groups using non-radioactive DNA-DNA dot-blot hybridisation. Rhizobium leguminosarum, R. galegae, R. tropici, Mesorhizobium loti, Sinorhizobium fredii, S. meliloti used in numerical taxonomy were included in the hybridisation experiments as reference strains and, at a later stage S. saheli and S. terangae . Scores given to the intensities of dots detected in the hybridisation experiments were used in principal component analysis, which clustered the majority of the tree rhizobia in two separate DNA-homology groups. The 51 strains were also analysed by genomic fingerprinting using the repetitive sequence-based polymerase chain reaction (rep-PCR) method with REP, ERIC, BOX and GTG5 primers. The resulting genomic fingerprints were compared with strains representing 15 rhizobial species. The relationship of 17 Sudanese strains to established sinorhizobial species was examined using the optical renaturation rates method and the G+C content of nine strains was determined. Results from dot-blot hybridisation and rep-PCR experiments were found to be in close agreement with each other and with the results obtained from spectrophotometric reassociation analysis. We suggest that rep-PCR fingerprinting can be used as a first and dot-blot hybridisation as a second rapid and dependable genomic screening method to classify new rhizobial isolates of unknown taxonomic status and to choose the representative strains for the more laborious DNA-DNA reassociation experiments.

Histopathological Study of Three Leishmania Major Strains

Amastigotes count in inoculated foot pad and histopathologicaJ study of infected mice were performed to study three Leishmania major strains: the Senegalese strain MHOM/SN/OOIDKI(SN), the Sudanese strain MHOMlSO/89/UG8(SO), and the Saudi strain MHOMlSAl84/JISH(SA). Eight-week-old male BALB/c mice were divided into three groups. Each group was infected with one of the studied strains. Amastigotes in the inoculat- ed foot of infected.mice in the three groups were counted at 30 and 45 days us- ing modified fluorescence diacetate/ethidium bromide viability test, The num- ber of amastigotes in the SA group was found to be significantly lower than that of the other two groups at both durations. There was no significant differ- ence between the number of amastigotes in the SN and SO strains at 30 days, but it was significantly higher in the SN group than that of the SD group at 45 days post-inoculation. In the animals studied, tissue samples were obtained from the skin, liver, spleen, and testes for gross pathological changes and histo- pathological evaluation. The SO strain started earlier and led a more aggres- sive course. The SN and SA strains were less aggressive. The SA strain showed changes at an earlier time and resulted in a more aggressive course of the disease. The SN and SA strains were less aggressive. The SA strain showed the most chronic course of the three.

Serogrouping and topotyping of Sudanese and United States strains of epizootic hemorrhagic disease virus using PCR

The potential use of the recently reported polymerase chain reaction (PCR) protocol for detection of United States epizootic hemorrhagic disease virus (EHDV) serotype 1 (EHDV-1) and serotype 2 (EHDV-2) ribonucleic acid in cell culture and clinical specimens was evaluated for detection of Sudanese EHDV strains. EHDV serotype 5 (EHDV-5) and EHDV, isolate 318 (untyped) designated (EHDV-318), recovered from sentinel calves at the Khartoum University farm (Sudan) were studied. RNA from EHDV-5 and EHDV-318 and a number of EHDV field isolates, propagated in cell cultures, were detected by the described PCR-based assay. The specific 387 bp PCR products were visualized on ethidium-bromide stained agarose gel. Specificity of the PCR products was confirmed by chemiluminescent hybridization with non-radiolabeled internal probe. Amplification product was not detected when the PCR-based assay was applied to RNA from bluetongue virus (BTV) prototypes serotypes 2, 10, 11, 13, 16 and 17; total nucleic acid extracts from uninfected BHK-21 cells. The results of this study indicated that the previously described EHDV-PCR assay could be applied for detection of Sudanese as well as United States strains of EHDV serogroup. In addition, the described EHDV-PCR assay could be used as a supportive diagnostic assay to the current conventional virus isolation procedures used for detection of EHDV infection in susceptible ruminants.