Avtor/Urednik		Gutierrez-Aguirre, Ion; Steyer, Andrej; Boben, Jana; Gruden, Kristina; Poljšak-Prijatelj, Mateja; Ravnikar, Maja
Naslov		Sensitive detection of multiple rotavirus genotypes with a single RT-qPCR assay
Tip		članek
Vir		J Clin Microbiol
Vol. in št.		Letnik 48, št. 8
Leto izdaje		2008
Obseg		str. 2547-54
Jezik		eng
Abstrakt		Rotaviruses are one of the major causes of diarrhea in infants and children under 5 years old, affecting specially developing countries. In case of natural disasters, fecal and potable waters can mix, allowing low, yet infective, concentrations of rotavirus to be present in water supplies, thus, constituting a risk for the population. Any of the most commonly detected rotavirus genotypes could originate an outbreak. The development of a fast and sensitive method which could detect the broadest possible range of rotavirus genotypes would help on efficient diagnosis and prevention. We have designed a reverse transcription-real time quantitative polymerase chain reaction (RT-qPCR) approach targeted to the rotaviral VP2 gene, based on a multi-sequence alignment of different human rotaviral strains. To overcome the high nucleotide sequence diversity multiple forward and reverse primers were used in addition to a degenerated probe. The performance of the assay was tested on isolates representing the most prevalent human genotypes: G1P[8], G2P[4], G3P[8], G4P[8], G9P[8], and G12P[8]. The developed method improved classical rotavirus detection by ELISA and nested RT-PCR by five and, at least one order of magnitude, respectively. A survey of 159 stool samples indicated that the developed method can efficiently detect a broad range of rotavirus strains, including different G-P genotype combinations from human, porcine and bovine origin. No cross reactivity was observed with other enteric viruses, such as, Astrovirus, Sapovirus and Norovirus.
Deskriptorji		ROTAVIRUS ROTAVIRUS INFECTIONS DIARRHEA GENOTYPE POLYMERASE CHAIN REACTION INFANT CHILD, PRESCHOOL