Development of a rapid immunochromatographic test for noroviruses genogroups I and II.
Norovirus (NoV) is known to cause acute gastroenteritis in children worldwide. Although reverse transcription-PCR (RT-PCR) method is considered to be the "gold standard" for diagnosis of this viral infection, it requires skillful personnel and well-equipped laboratory. In this study, a rapid and easily performable diagnostic kit was developed using immunochromatographic method with rabbit polyclonal antibodies raised against recombinant virus-like particles (rVLPs) of most prevalent genotypes, genogroup II genotypes 3 and 4. This kit was evaluated for reactivity to rVLPs and detection of natural viruses in stool samples collected from children with diarrhea in comparison to the results obtained by RT-PCR. In the prospective assessment, the kit showed agreement rate of 84.1%, sensitivity of 69.8% and specificity of 93.7%. Genotyping of the RT-PCR positive samples by sequence analysis revealed that some heterogeneous genotypes were also detected while some in homogeneous genotypes occasionally showed false negative records resulting in lower sensitivity. No cross-reactivity with other common viral pathogens was observed. Taken together with the result of the detection limit of viral load as small as approximately 10(6-7)copies/g of stool, the current immunochromatography test is justified for screening for NoV infection with simple laboratory support.
Takanashi
Sayaka
S
Department of Developmental Medical Sciences, Graduate School of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
Okame
Michio
M
Shiota
Tomoyuki
T
Takagi
Makiko
M
Yagyu
Fumihiro
F
Tung
Phan Gia
PG
Nishimura
Syuichi
S
Katsumata
Noriko
N
Igarashi
Takashi
T
Okitsu
Shoko
S
Ushijima
Hiroshi
H
D000818:Animals; D000914:Antibodies, Viral; D017250:Caliciviridae Infections / Q000175:diagnosis* / Q000821:virology; D002648:Child; D002845:Chromatography / Q000379:methods*; D003429:Cross Reactions; D005188:False Negative Reactions; D005243:Feces / Q000821:virology; D005759:Gastroenteritis / Q000175:diagnosis / Q000821:virology*; D005838:Genotype; D006801:Humans; D029322:Norovirus / Q000145:classification* / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D010802:Phylogeny; D012367:RNA, Viral / Q000235:genetics; D011817:Rabbits; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D022701:Virosomes / Q000276:immunology
D000914:Antibodies, Viral; D012367:RNA, Viral; D022701:Virosomes