Multiplex polymerase chain reaction amplification and differentiation of Entamoeba histolytica and Entamoeba dispar DNA from stool samples

被引:40
作者
Núñez, YO
Fernández, MA
Torres-Núñez, D
Silva, JA
Montano, I
Maestre, JL
Fonte, L
机构
[1] Ctr Med & Surg Res, Havana, Cuba
[2] Ctr Genet Engn & Biotechnol, Havana, Cuba
[3] Inst Trop Med Pedro Kouri, Havana, Cuba
关键词
D O I
10.4269/ajtmh.2001.64.293
中图分类号
R1 [预防医学、卫生学];
学科分类号
1004 ; 120402 ;
摘要
Due to the clinical importance of differentiating the two species of the Entamoeba histolytica/Entamoeba dispar complex, we developed a multiplex polymerase chain reaction (PCR) method that overcomes time-consuming and laborious procedures. We report here a DNA extraction protocol using non-fixed stool samples that avoid long lysis-incubation periods through the combined use of zirconium beads and a lysis-supporting buffer. We characterized 49 of 52 stool specimens from Cuban patients with amoebiosis. Among them, 36 (75.5%) were infected only with E. dispar (the nonpathogenic species), while 13 (24.5%) displayed a mixed infection with both E. dispar and E. histolytica. The multiplex PCR protocol showed a specificity of 1.00 and a sensitivity of 0.94. Furthermore, the entire procedure can be performed in one day. This approach is therefore reliable and applicable in the field for epidemiologic studies.
引用
收藏
页码:293 / 297
页数:5
相关论文
共 44 条
  • [1] Acuna-Soto R, 2000, AM J GASTROENTEROL, V95, P1277, DOI 10.1111/j.1572-0241.2000.01525.x
  • [2] APPLICATION OF THE POLYMERASE CHAIN-REACTION TO THE EPIDEMIOLOGY OF PATHOGENIC AND NONPATHOGENIC ENTAMOEBA-HISTOLYTICA
    ACUNASOTO, R
    SAMUELSON, J
    DEGIROLAMI, P
    ZARATE, L
    MILLANVELASCO, F
    SCHOOLNICK, G
    WIRTH, D
    [J]. AMERICAN JOURNAL OF TROPICAL MEDICINE AND HYGIENE, 1993, 48 (01) : 58 - 70
  • [3] AMPLIFICATION OF NUCLEIC-ACIDS BY POLYMERASE CHAIN-REACTION (PCR) AND OTHER METHODS AND THEIR APPLICATIONS
    BEJ, AK
    MAHBUBANI, MH
    ATLAS, RM
    [J]. CRITICAL REVIEWS IN BIOCHEMISTRY AND MOLECULAR BIOLOGY, 1991, 26 (3-4) : 301 - 334
  • [4] MULTIPLEX PCR AMPLIFICATION AND IMMOBILIZED CAPTURE PROBES FOR DETECTION OF BACTERIAL PATHOGENS AND INDICATORS IN WATER
    BEJ, AK
    MAHBUBANI, MH
    MILLER, R
    DICESARE, JL
    HAFF, L
    ATLAS, RM
    [J]. MOLECULAR AND CELLULAR PROBES, 1990, 4 (05) : 353 - 365
  • [5] DIFFERENTIATION OF CLINICAL ISOLATES OF ENTAMOEBA-HISTOLYTICA BY USING SPECIFIC DNA PROBES
    BRACHA, R
    DIAMOND, LS
    ACKERS, JP
    BURCHARD, GD
    MIRELMAN, D
    [J]. JOURNAL OF CLINICAL MICROBIOLOGY, 1990, 28 (04) : 680 - 684
  • [6] SEROEPIDEMIOLOGY OF AMEBIASIS IN MEXICO
    CABALLEROSALCEDO, A
    VIVEROSROGEL, M
    SALVATIERRA, B
    TAPIACONYER, R
    SEPULVEDAAMOR, J
    GUTIERREZ, G
    ORTIZORTIZ, L
    [J]. AMERICAN JOURNAL OF TROPICAL MEDICINE AND HYGIENE, 1994, 50 (04) : 412 - 419
  • [7] DELETION SCREENING OF THE DUCHENNE MUSCULAR-DYSTROPHY LOCUS VIA MULTIPLEX DNA AMPLIFICATION
    CHAMBERLAIN, JS
    GIBBS, RA
    RANIER, JE
    NGUYEN, PN
    CASKEY, CT
    [J]. NUCLEIC ACIDS RESEARCH, 1988, 16 (23) : 11141 - 11156
  • [8] Chamberlain JS, 1990, PCR PROTOCOLS GUIDE, P272
  • [9] ENTAMOEBA-HISTOLYTICA - AN EXPLANATION FOR THE REPORTED CONVERSION OF NONPATHOGENIC AMEBAE TO THE PATHOGENIC FORM
    CLARK, CG
    DIAMOND, LS
    [J]. EXPERIMENTAL PARASITOLOGY, 1993, 77 (04) : 456 - 460
  • [10] RIBOSOMAL DNA-SEQUENCES IN THE DIFFERENTIATION OF PATHOGENIC AND NONPATHOGENIC ISOLATES OF ENTAMOEBA-HISTOLYTICA
    CRUZREYES, JA
    SPICE, WM
    REHMAN, T
    GISBORNE, E
    ACKERS, JP
    [J]. PARASITOLOGY, 1992, 104 : 239 - 246