scholarly journals Simultaneous detection of fourteen respiratory viruses in clinical specimens by two multiplex reverse transcription nested-PCR assays

2004 ◽  
Vol 72 (3) ◽  
pp. 484-495 ◽  
Author(s):  
M.T. Coiras ◽  
J.C. Aguilar ◽  
M.L. García ◽  
I. Casas ◽  
P. Pérez-Breña
Keyword(s):  
Reverse Transcription ◽  
Nested Pcr ◽  
Simultaneous Detection ◽  
Respiratory Viruses ◽  
Clinical Specimens ◽  
Pcr Assays ◽  
Nested Pcr Assays

scholarly journals Duplex Reverse Transcription-PCR Followed by Nested PCR Assays for Detection and Identification of Brazilian Alphaviruses and Flaviviruses

2005 ◽  
Vol 43 (2) ◽  
pp. 696-702 ◽  
Author(s):  
R. V. d. M. Bronzoni ◽  
F. G. Baleotti ◽  
R. M. Ribeiro Nogueira ◽  
M. Nunes ◽  
L. T. Moraes Figueiredo
Keyword(s):  
Reverse Transcription ◽  
Nested Pcr ◽  
Reverse Transcription Pcr ◽  
Detection And Identification ◽  
Pcr Assays ◽  
Nested Pcr Assays

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

2004 ◽  
Vol 70 (3) ◽  
pp. 1448-1454 ◽  
Author(s):  
Carlos Maluquer de Motes ◽  
Pilar Clemente-Casares ◽  
Ayalkibet Hundesa ◽  
Margarita Mart�n ◽  
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.

scholarly journals Rapid and Sensitive Detection of Noroviruses by Using TaqMan-Based One-Step Reverse Transcription-PCR Assays and Application to Naturally Contaminated Shellfish Samples

2005 ◽  
Vol 71 (4) ◽  
pp. 1870-1875 ◽  
Author(s):  
Narayanan Jothikumar ◽  
James A. Lowther ◽  
Kathleen Henshilwood ◽  
David N. Lees ◽  
Vincent R. Hill ◽  
...  
Keyword(s):  
Reverse Transcription ◽  
Nested Pcr ◽  
Rt Pcr ◽  
Reverse Transcription Pcr ◽  
Specificity And Sensitivity ◽  
Routine Monitoring ◽  
The Family ◽  
One Step ◽  
Stool Specimens ◽  
Pcr Assays

ABSTRACT Noroviruses (NoV), which are members of the family Caliciviridae, are the most important cause of outbreaks of acute gastroenteritis worldwide and are commonly found in shellfish grown in polluted waters. In the present study, we developed broadly reactive one-step TaqMan reverse transcription (RT)-PCR assays for the detection of genogroup I (GI) and GII NoV in fecal samples, as well as shellfish samples. The specificity and sensitivity of all steps of the assays were systematically evaluated, and in the final format, the monoplex assays were validated by using RNA extracted from a panel of 84 stool specimens, which included NoV strains representing 19 different genotypes (7 GI, 11 GII, and 1 GIV strains). The assays were further validated with 38 shellfish cDNA extracts previously tested by nested PCR. Comparison with a recently described real-time assay showed that our assay had significantly higher sensitivity and was at least as sensitive as the nested PCR. For stool specimens, a one-step duplex TaqMan RT-PCR assay performed as well as individual genogroup-specific monoplex assays. All other enteric viruses examined were negative, and no cross-reaction between genogroups was observed. These TaqMan RT-PCR assays provide rapid (less than 90 min), sensitive, and reliable detection of NoV and should prove to be useful for routine monitoring of both clinical and shellfish samples.

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

1996 ◽  
Vol 62 (2) ◽  
pp. 131-141 ◽  
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

scholarly journals 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.

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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