Publication: Rapid detection and identification of dengue viruses by polymerase chain reaction (PCR)
Issued Date
1996-06-01
Resource Type
ISSN
01251562
Other identifier(s)
2-s2.0-0030154667
Rights
Mahidol University
Rights Holder(s)
SCOPUS
Bibliographic Citation
Southeast Asian Journal of Tropical Medicine and Public Health. Vol.27, No.2 (1996), 228-236
Suggested Citation
Pa Thai Yenchitsomanus, Pilaichit Sricharoen, Isaya Jaruthasana, Sa Nga Pattanakitsakul, Sorachai Nitayaphan, Juthathip Mongkolsapaya, Prida Malasit Rapid detection and identification of dengue viruses by polymerase chain reaction (PCR). Southeast Asian Journal of Tropical Medicine and Public Health. Vol.27, No.2 (1996), 228-236. Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/17740
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Title
Rapid detection and identification of dengue viruses by polymerase chain reaction (PCR)
Abstract
A polymerase chain reaction (PCR) method using sets of newly designed primers for rapid detection and simultaneous identification of dengue virus serotypes was developed and tested. The test is based on two sets of primers specific within the envelope (E) and non-structural (NS1) regions of the dengue-virus genome. Two sets of universal primers that bind to two target sequences which are shared by all the four serotypes of the virus within the E and NS1 regions are used. The resulting products are further amplified by another pair of inner or nested universal primers, which also bind to another set of shared sequences within the E and NS1 regions, respectively. The nested PCR of both the E and NS1 regions can detect dengue virus of all the four serotypes at a sensitivity of 1 plaque forming unit (pfu) or less. For the identification of serotypes, a mixture of four pairs of serotype-specific primers, specific to the E region, was used. The primers have been designed to bind to serotype specific sequences within the regions flanked by the outer universal primers, and giving the amplified products of different sizes, each corresponds to one particular serotype (405 bp for Den1, 346 bp for Den2, 196 bp for Den3, and 143 bp for Den4). A protocol has been developed and successfully applied to detect dengue virus in cell-culture supernatants and patients sera. The technique is simple and rapid, capable of not only detecting the dengue virus but also identifying the serotypes of the virus in clinical specimens.