Rapid diagnosis of adenoviral conjunctivitis by PCR and restriction fragment length polymorphism analysis

被引:91
作者
SaitohInagawa, W
Oshima, A
Aoki, K
Itoh, N
Isobe, K
Uchio, E
Ohno, S
Nakajima, H
Hata, K
Ishiko, H
机构
[1] MITSUBISHI KAGAKU BIOCLIN LABS INC,ITABASHI KU,TOKYO 174,JAPAN
[2] AOKI EYE CLIN,SHIROISHI KU,SAPPORO,HOKKAIDO,JAPAN
关键词
D O I
10.1128/JCM.34.9.2113-2116.1996
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
To detect and identify adenovirus (Ad), we used a combination of PCR and restriction fragment length polymorphism (RFLP) analysis. Nested PCR with two primer sets that hybridize to the conserved region for hexon proteins of 14 prototypes of Ad, Ad serotype 1 (Ad1) to Ads, -11, -14, -19, -37, -40, and -41, amplified a 956-bp DNA fragment. The amplified fragments from the 14 prototypes were completely differentiated with a combination of three restriction endonucleases, EcoT14I, HaeIII, and HinfI. We applied this new method for 127 samples of conjunctival scrapings from patients with conjunctivitis and compared the results with those obtained with the combination of culture isolation and a neutralization test (NT). PCR gave a positive result in 69 of 127 cases (54.3%), while only 61 of the 127 samples (48.0%) tested positive by culture isolation. Compared with isolation, the PCR method had a sensitivity of 100% (61 of 61). Positive PCR samples were further classified as Ad37 (59.5%), -3 (31.9%), -11 (4.3%), -8 (2.9%), and -4 (1.4%) by PCR-RFLP analysis. Of eight samples that were PCR positive and culture isolation negative, six were Ad37 and two were Ads by PCR-RFLP analysis. These differentiations of isolation-positive samples were identical to the results obtained by the NT. It took only 3 days to detect and identify Ad by PCR-RFLP analysis, whereas it took at least 3 weeks by culture isolation and NT. Our newly developed method of detecting and typing human Ad by PCR-RFLP analysis is more sensitive, accurate, and rapid than the conventional method of culture isolation and an NT.
引用
收藏
页码:2113 / 2116
页数:4
相关论文
共 15 条
  • [1] POLYMERASE CHAIN-REACTION FOR DETECTION OF ADENOVIRUSES IN STOOL SAMPLES
    ALLARD, A
    GIRONES, R
    JUTO, P
    WADELL, G
    [J]. JOURNAL OF CLINICAL MICROBIOLOGY, 1990, 28 (12) : 2659 - 2667
  • [2] CLINICAL AND ETIOLOGICAL STUDY OF ADENOVIRAL CONJUNCTIVITIS, WITH SPECIAL REFERENCE TO ADENOVIRUS TYPE-4 AND TYPE-19 INFECTIONS
    AOKI, K
    KATO, M
    OHTSUKA, H
    ISHII, K
    NAKAZONO, N
    SAWADA, H
    [J]. BRITISH JOURNAL OF OPHTHALMOLOGY, 1982, 66 (12) : 776 - 780
  • [3] AOKI K, 1986, JPN J OPHTHALMOL, V30, P158
  • [4] AOKI K, 1994, INFECT DIS EYE, P309
  • [5] THE SEQUENCE OF THE GENOME OF ADENOVIRUS TYPE-5 AND ITS COMPARISON WITH THE GENOME OF ADENOVIRUS TYPE-2
    CHROBOCZEK, J
    BIEBER, F
    JACROT, B
    [J]. VIROLOGY, 1992, 186 (01) : 280 - 285
  • [6] DETECTION OF ADENOVIRUS IN CLINICAL SPECIMENS BY POLYMERASE CHAIN-REACTION AND LIQUID-PHASE HYBRIDIZATION QUANTITATED BY TIME-RESOLVED FLUOROMETRY
    HIERHOLZER, JC
    HALONEN, PE
    DAHLEN, PO
    BINGHAM, PG
    MCDONOUGH, MM
    [J]. JOURNAL OF CLINICAL MICROBIOLOGY, 1993, 31 (07) : 1886 - 1891
  • [7] KINCHINGTON PR, 1994, INVEST OPHTH VIS SCI, V35, P4126
  • [8] NAKAGAWA H, 1995, J JPN OPHTHALMOL, V99, P1164
  • [9] PHILIPSON L, 1984, CURR TOP MICROBIOL I, V110, P1
  • [10] TYPE-SPECIFIC AND GROUP-SPECIFIC POLYMERASE CHAIN-REACTION FOR ADENOVIRUS DETECTION
    PRINGAKERBLOM, P
    ADRIAN, T
    [J]. RESEARCH IN VIROLOGY, 1994, 145 (01): : 25 - 35