Detection and identification of Toscana and other phleboviruses by RT-nested-PCR assays with degenerated primers

ABSTRACT In this study, a molecular procedure for the detection of adenoviruses of animal origin was developed to evaluate the level of excretion of these viruses by swine and cattle and to design a test to facilitate the tracing of specific sources of environmental viral contamination. Two sets of oligonucleotides were designed, one to detect porcine adenoviruses and the other to detect bovine and ovine adenoviruses. The specificity of the assays was assessed in 31 fecal samples and 12 sewage samples that were collected monthly during a 1-year period. The data also provided information on the environmental prevalence of animal adenoviruses. Porcine adenoviruses were detected in 17 of 24 (70%) pools of swine samples studied, with most isolates being closely related to serotype 3. Bovine adenoviruses were present in 6 of 8 (75%) pools studied, with strains belonging to the genera Mastadenovirus and Atadenovirus and being similar to bovine adenoviruses of types 2, 4, and 7. These sets of primers produced negative results in nested PCR assays when human adenovirus controls and urban-sewage samples were tested. Likewise, the sets of primers previously designed for detection of human adenovirus also produced negative results with animal adenoviruses. These results indicate the importance of further studies to evaluate the usefulness of these tests to trace the source of fecal contamination in water and food and for environmental studies.

Download Full-text

Nested PCR assays with novel primers yield greater sensitivity to Tanzanian HIV-1 samples than a commercial PCR detection kit

Journal of Virological Methods ◽

10.1016/s0166-0934(96)02094-0 ◽

1996 ◽

Vol 62 (2) ◽

pp. 131-141 ◽

Cited By ~ 4

Author(s):

Olov Grankvist ◽

Lilian Walther ◽

Ulla Bredberg-Rådén ◽

Eligius Lyamuya ◽

Fred Mhalu ◽

...

Keyword(s):

Nested Pcr ◽

Pcr Detection ◽

Pcr Assays ◽

Nested Pcr Assays ◽

Hiv 1

Download Full-text

Malaria Parasites in Macaques in Thailand: Stump-Tailed Macaques (Macaca Arctoides) are New Natural Hosts for Plasmodium Knowlesi, P. Inui, P. Coatneyi and P. Fieldi.

10.21203/rs.3.rs-26507/v1 ◽

2020 ◽

Author(s):

Wirasak Fungfuang ◽

Chanya Udom ◽

Daraka Tongthainan ◽

Khamisah Abdul Kadir ◽

Balbir Singh

Keyword(s):

Nested Pcr ◽

Plasmodium Knowlesi ◽

Sampling Site ◽

Macaca Arctoides ◽

Malaria Parasites ◽

Zoonotic Malaria ◽

Natural Hosts ◽

New Locations ◽

Pcr Assays ◽

Nested Pcr Assays

Abstract Background:Certain species of macaques are natural hosts ofPlasmodium knowlesi and P. cynomolgi, which can both cause malaria in humans, and P. inui, which can be experimentally transmitted to humans. A significant number of zoonotic malaria cases have been reported in humans throughout Southeast Asia, including Thailand. There have been only two studies undertaken in Thailand to identify malaria parasites in non-human primates in 6 provinces. The objective of this study was to determine the prevalence of P. knowlesi, P. coatneyi, P. cynomolgi, P. inui and P. fieldiin non-human primates from 4 new locations in Thailand. Methods:A total of 93 blood samples from Macaca fascicularis, M. leonina and M. arctoides were collected from four locations in Thailand: 32 were captive M. fascicularisfrom Chachoengsao Province (CHA), 4 were wild M. fascicularis from Ranong Province (RAN), 32 were wildM. arctoidesfromPrachuap Kiri Khan Province (PRA), and 25 were wild M. leoninafrom Nakornratchasima Province (NAK). DNA was extracted from these samples and analysed by nested PCR assays to detect Plasmodium, and subsequently to detectP. knowlesi, P. coatneyi, P. cynomolgi, P. inui and P. fieldi.Results:Twenty-seven of the 93 (29%) samples were Plasmodium-positive by nested PCR assays. Among wild macaques, all 4 M. fascicularis at RAN were infected with malaria parasites followed by 50% of 32 M. arctoides at PRA and 20% of 25 M. leonina at NAK. Only 2 (6.3%) of the 32 captive M. fascicularisat CHA were malaria-positive. All 5 species of Plasmodium were detected and 16 (59.3%) of the 27 macaques had single infections, 9had double and 2 had triple infections.The composition of Plasmodium species in macaques at each sampling site was different. Macaca arctoides from PRA were infected with P. knowlesi, P. coatneyi, P. cynomolgi, P. inui and P. fieldi. Conclusions:The prevalence and species of Plasmodiumvaried among the wild and captive macaques, and betweenmacaques at 4 sampling sites in Thailand. Macaca arctoidesis a new natural host for P. knowlesi, P. inui,P. coatneyi and P. fieldi.

Download Full-text

Molecular Detection and Identification of Babesia spp., and Trypanosoma spp. in One-humped Camel (Camelus dromedarius) in Halayeb and Shalateen, Egypt

10.20944/preprints202010.0334.v1 ◽

2020 ◽

Author(s):

Shimaa Abd El-Salam El-Sayed ◽

Mohamed El-Adl ◽

Mayar Ali ◽

Mostafa Al-Araby ◽

Mosaab Omar ◽

...

Keyword(s):

Infection Rate ◽

Phylogenetic Analyses ◽

Small Subunit ◽

Camelus Dromedarius ◽

Babesia Microti ◽

Small Subunit Rrna ◽

Detection And Identification ◽

Pcr Assays ◽

Nested Pcr Assays ◽

Babesia Spp

Phylogenetic analysis of blood parasite infections including Babesia (B.) bovis, Babesia microti and Trypanosoma (T.) spp. in one-humped camel (Camelus dromedarius) (n= 142) breeds in Halayeb and Shalateen, in Upper Egypt were performed in the current study. Polymerase chain reaction (PCR) assays targeting the Rhoptry Associated Protein-1 (RAP-1), Babesia microti small subunit rRNA (ss-rRNA) and internal transcribed spacer 1 (ITS1) genes were used to detect the prevalence of B. bovis, B. microti and Trypanosoma spp. in camels, respectively. Nested PCR assays were used for the detection of Babesia spp. (B. bovis and B. microti). While, KIN-multi species PCR reaction was employed to detect and identify trypanosome DNA in camels. B. microti was detected in (17/142) with infection rate (11.97 %). Sequencing and phylogenetic analyses revealed that B. microti detected in camel was closely related to the German strain in rats and voles in France. B. bovis was also detected in (4/142) with infection rate (2.81%). The sequence and phylogenetic analyses revealed that the isolated B. bovis was closely related to strains isolated from Argentine, USA and Brazil. Moreover, T. evansi was detected in (8/142) with infection rate (5.63%). Sequence and phylogenetic analyses revealed that isolated T. evansi was closely related to T. theileri that was detected from cattle in Brazil. This study provides the first evidence of B. microti in camel in Egypt and highlights the possible role of one-humped camels in maintaining the enzootic cycle of Babesia transmission in Egypt.

Download Full-text