DETECTION OF GROUP-B AND GROUP-C ROTAVIRUSES BY POLYMERASE CHAIN-REACTION

被引:151
作者
GOUVEA, V
ALLEN, JR
GLASS, RI
FANG, ZY
BREMONT, M
COHEN, J
MCCRAE, MA
SAIF, LJ
SINARACHATANANT, P
CAUL, EO
机构
[1] US FDA,WASHINGTON,DC 20204
[2] CHINESE ACAD PREVENT MED,INST VIROL,BEIJING,PEOPLES R CHINA
[3] LAB VIROL & IMMUNOL MOLEC,F-78352 JOUY EN JOSAS,FRANCE
[4] UNIV WARWICK,DEPT BIOL SCI,COVENTRY CV4 7AL,W MIDLANDS,ENGLAND
[5] PUBL HLTH LAB,REG VIRUS LAB,BRISTOL BS2 8EL,ENGLAND
[6] OHIO STATE UNIV,OHIO AGR RES & DEV CTR,WOOSTER,OH 44691
[7] MAHIDOL UNIV,FAC SCI,DEPT MICROBIOL,BANGKOK 10700,THAILAND
关键词
D O I
10.1128/JCM.29.3.519-523.1991
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
We adapted the polymerase chain reaction (PCR) to detect the noncultivatable group B and C rotaviruses and introduced a simple and convenient technique to purify viral RNA from stool specimens. Double-stranded RNA present in stool extracts was purified by adsorption to hydrodroxyapatite and was used as the template for reverse transcription and polymerase amplification. Primer pairs specific for group B (gene 8) and group C (gene 6) rotaviruses were selected to amplify group-characteristic sizes of cDNA copies readily identifiable in ethidium bromide-stained agarose gels. These primer pairs were used separately in individual PCR assays or were pooled with a primer pair specific for group A rotavirus (gene 9) in a combined PCR assay for the simultaneous detection of all three rotavirus groups. The method was very sensitive and was used to identify both human and porcine strains of group B and C rotaviruses in stool specimens. A second PCR amplification with internal group-specific primers served to increase further the sensitivity of the test and to confirm the diagnostic results obtained in the first amplification.
引用
收藏
页码:519 / 523
页数:5
相关论文
共 26 条
  • [1] Bernardi G., 1971, METH ENZYMOL, V21, P95
  • [2] BREMONT M, 1990, VIROLOGY, V178, P597
  • [3] GROUP-C ROTAVIRUSES IN HUMANS
    BRIDGER, JC
    PEDLEY, S
    MCCRAE, MA
    [J]. JOURNAL OF CLINICAL MICROBIOLOGY, 1986, 23 (04) : 760 - 763
  • [4] BROWN DW, 1990, 8TH INT C VIR, P434
  • [5] ROTAVIRUS EPIDEMIOLOGY IN VELLORE, SOUTH-INDIA - GROUP, SUBGROUP, SEROTYPE, AND ELECTROPHORETYPE
    BROWN, DWG
    MATHAN, MM
    MATHEW, M
    MARTIN, R
    BEARDS, GM
    MATHAN, VI
    [J]. JOURNAL OF CLINICAL MICROBIOLOGY, 1988, 26 (11) : 2410 - 2414
  • [6] PREVALENCE OF ANTIBODY TO GROUP-B (ATYPICAL) ROTAVIRUS IN HUMANS AND ANIMALS
    BROWN, DWG
    BEARDS, GM
    CHEN, GM
    FLEWETT, TH
    [J]. JOURNAL OF CLINICAL MICROBIOLOGY, 1987, 25 (02) : 316 - 319
  • [7] CHARACTERIZATION OF MONOCLONAL-ANTIBODIES TO HUMAN GROUP-B ROTAVIRUS AND THEIR USE IN AN ANTIGEN-DETECTION ENZYME-LINKED IMMUNOSORBENT-ASSAY
    BURNS, JW
    WELCH, SKW
    NAKATA, S
    ESTES, MK
    [J]. JOURNAL OF CLINICAL MICROBIOLOGY, 1989, 27 (02) : 245 - 250
  • [8] GROUP-C ROTAVIRUS ASSOCIATED WITH FATAL ENTERITIS IN A FAMILY OUTBREAK
    CAUL, EO
    ASHLEY, CR
    DARVILLE, JM
    BRIDGER, JC
    [J]. JOURNAL OF MEDICAL VIROLOGY, 1990, 30 (03) : 201 - 205
  • [9] CDNA CLONING OF EACH GENOMIC SEGMENT OF THE GROUP-B ROTAVIRUS ADRV - MOLECULAR CHARACTERIZATION OF THE 11TH RNA SEGMENT
    CHEN, GM
    HUNG, T
    MACKOW, ER
    [J]. VIROLOGY, 1990, 175 (02) : 605 - 609
  • [10] PURIFICATION AND CHARACTERIZATION OF ADULT DIARRHEA ROTAVIRUS - IDENTIFICATION OF VIRAL STRUCTURAL PROTEINS
    FANG, ZY
    GLASS, RI
    PENARANDA, M
    DONG, H
    MONROE, SS
    WEN, L
    ESTES, MK
    EIDEN, J
    YOLKEN, RH
    SAIF, L
    GOUVEA, V
    HUNG, T
    [J]. JOURNAL OF VIROLOGY, 1989, 63 (05) : 2191 - 2197