Detection and Genotyping of Francisella tularensis in Animal Hosts and Vectors from Six Different Natural Landscape Areas, Gansu Province, China

Objective. Tularemia, also known as hare fever, is caused by the bacterium Francisella tularensis (F. tularensis) transmitted through diseased wild animals, blood sucking insects, or contaminated water or food, which is distributed worldwide. The purpose of this study was to investigate F. tularensis infection in animal hosts and vectors from six different natural landscape areas in Gansu Province and to identify the genotypes of the detected F. tularensis. Methods. Rodents were captured by snap traps, and ticks were collected by dragging a cloth over the vegetation or from domestic animals. After species identification, DNA was isolated from the captured animals and detected by nested PCR assays targeting the F. tularensis fopA gene. The positive samples were further amplified to discriminate the species, and another two short-sequence tandem repeat regions (SSTR) were amplified to identify their genotypes. All positive fragments were sequenced and analyzed by ClustalX (5.0) and DNAClub software. Results. A total of 407 rodents of 12 species were captured, among which six rodent species were positive for F. tularensis, with an overall prevalence of 3.93%. The geographical difference in infection rate was statistically significant. At the SSTR9 locus, there were 7 genotypes among positive rodent samples. A total of 1864 ticks were tested for evidence of tularemia by nested PCR assays, 69 of which were positive, with an average positive rate of 3.70% for F. tularensis in ticks. The positive rates were significantly different among different regions. Seven genotypes were identified at the SSTR9 locus, one of which seemed dominant in positive tick samples. All positive samples had the same genotype at the SSTR16 locus. Conclusion. There is natural infection of F. tularensis among animal vectors and hosts in Gansu Province, with diverse genotypes.

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Development of Nested PCR Assays for Detection of Bovine Respiratory Syncytial Virus in Clinical Samples

Journal of Clinical Microbiology ◽

10.1128/jcm.32.11.2887-2887.1994 ◽

1994 ◽

Vol 32 (11) ◽

pp. 2887-2887

Keyword(s):

Respiratory Syncytial Virus ◽

Nested Pcr ◽

Bovine Respiratory Syncytial Virus ◽

Clinical Samples ◽

Pcr Assays ◽

Syncytial Virus ◽

Nested Pcr Assays

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Detection of Bovine and Porcine Adenoviruses for Tracing the Source of Fecal Contamination

Applied and Environmental Microbiology ◽

10.1128/aem.70.3.1448-1454.2004 ◽

2004 ◽

Vol 70 (3) ◽

pp. 1448-1454 ◽

Cited By ~ 70

Author(s):

Carlos Maluquer de Motes ◽

Pilar Clemente-Casares ◽

Ayalkibet Hundesa ◽

Margarita MartÃ¯Â¿Â½n ◽

Rosina Girones

Keyword(s):

Nested Pcr ◽

Fecal Contamination ◽

Human Adenovirus ◽

The Other ◽

Animal Origin ◽

Viral Contamination ◽

Urban Sewage ◽

Negative Results ◽

Pcr Assays ◽

Nested Pcr Assays

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.

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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

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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.

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