1993
DOI: 10.1128/jcm.31.11.3028-3030.1993
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Sensitive identification of bluetongue virus serogroup by a colorimetric dual oligonucleotide sorbent assay of amplified viral nucleic acid

Abstract: A polymerase chain reaction (PCR) procedure coupled to an enzyme-linked oligonucleotide sorbent assay (ELOSA; a PCR-ELOSA) identified all 24 serotypic variants of bluetongue virus (BTV) without identifying any of six viruses belonging to the related epizootic hemorrhagic disease virus serogroup. The PCR-ELOSA detected 0.01 50%o cell culture infectious doses of each serotype of BTV. The sensitivity and serogroup-wide specificity of the PCR-ELOSA may enable it to replace the more expensive, time-consuming, and b… Show more

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Cited by 45 publications
(12 citation statements)
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“…All PCRs were carried out with denaturation at 94°C for 15 s, annealing at 55°C for 30 s and extension at 68°C with one minute for every thousand bases of the PCR product. The isolate was confirmed as BTV by Seg‐5‐ and Seg‐7‐based conventional RT‐PCR as described earlier (Katz et al., ; OIE, , Wade‐Evans et al., ). Initially, IND2010/01 cDNA was subjected to serotype‐specific RT‐PCR for eight common serotypes identified during the last decade from India (Reddy et al., ), followed by primers against 26 global serotypes of BTV (Maan et al., ) to identify the serotype of the isolate.…”
Section: Methodsmentioning
confidence: 90%
“…All PCRs were carried out with denaturation at 94°C for 15 s, annealing at 55°C for 30 s and extension at 68°C with one minute for every thousand bases of the PCR product. The isolate was confirmed as BTV by Seg‐5‐ and Seg‐7‐based conventional RT‐PCR as described earlier (Katz et al., ; OIE, , Wade‐Evans et al., ). Initially, IND2010/01 cDNA was subjected to serotype‐specific RT‐PCR for eight common serotypes identified during the last decade from India (Reddy et al., ), followed by primers against 26 global serotypes of BTV (Maan et al., ) to identify the serotype of the isolate.…”
Section: Methodsmentioning
confidence: 90%
“…Previously published primers were used for NS1 and VP7 genes (Wade‐Evans et al., 1990; Katz et al., 1993; OIE, 2010), and for VP2 , NS2 and VP6 genes, primers were designed using VP2, NS2 and VP6 sequences of USA BTV‐10 vaccine virus using NCBI‐Primer Blast (Rozen and Skaletsky, 2000) program (Table 1). Infected cell lysate was subjected to centrifugation, and RNA was extracted from pelleted cells and membranes using Trizol‐LS ® (Invitrogen Bioservices Private Limited, Bangalore, India), denatured at 65°C for 10 min and snap cooled on ice.…”
Section: Methodsmentioning
confidence: 99%
“…Recently, several investigators have presented sensitive assays for the diagnosis of various viral (16,17,24,30,35), bacterial (3,5,7,8,20,22,23,28,33,37), fungal (14), and protozoal (2) infections on the basis of nonradioactive (colorimetric or chemiluminescent) nucleic acid hybridization. These kinds of techniques are simple, and colorimetry requires only a conventional microtiter spectrophotometric reader and can be readily done on a large scale.…”
mentioning
confidence: 99%