Neospora caninum: Specific oligonucleotide primers for the detection of brain ''cyst'' DNA of experimentally infected nude mice by the polymerase chain reaction (PCR)

被引:237
作者
Yamage, M [1 ]
Flechtner, O [1 ]
Gottstein, B [1 ]
机构
[1] UNIV BERN,FAC MED,BERN,SWITZERLAND
关键词
D O I
10.2307/3284160
中图分类号
R38 [医学寄生虫学]; Q [生物科学];
学科分类号
07 ; 0710 ; 09 ; 100103 ;
摘要
Both tachyzoites and bradyzoites in tissue cysts of Neospora caninum are morphologically difficult to distinguish from those of other cyst-forming apicomplexan parasites such as Toxoplasma gondii. Thus, molecular tools may contribute to easy identification of the parasite. Based upon an N. caninum-specific DNA fragment that we have recently cloned, 5 sense (Npl, Np3, Np5, Np7, Np21) and 4 antisense (Np2, Np4, Np5, Np8) oligonucleotides were designed for a sensitive and specific polymerase chain reaction (PCR). Among 19 combinations of sense and antisense primers, the Np21/Np6, Np21/Np4, and Np7/Np6 primer pairs were found to generate specific single bands in the presence of at least 10 pg genomic parasite DNA as a template. The primer pair Np21/Np6 was able to detect a single tachyzoite in the background of DNA derived from 2 mg of blain tissue. In experimentally infected athymic ICR:nu/nu mice, N. caninum- DNA was detected consistently from brain tissue at days 13 and 18 after subcutaneous inoculation of tachyzoites. The presence or absence of the organisms in the cerebrum in either proliferative or cystic form was examined by immunohistological staining. The results indicate that PCR with the primer pair Np21/Np6 could provide an efficient tool for large-scale epidemiological studies using brain tissue obtained at necropsy.
引用
收藏
页码:272 / 279
页数:8
相关论文
共 25 条
[11]  
DUBEY JP, 1988, J AM VET MED ASSOC, V192, P1269
[12]   NEOSPOROSIS [J].
DUBEY, JP ;
LINDSAY, DS .
PARASITOLOGY TODAY, 1993, 9 (12) :452-458
[13]   THE PHYLOGENY OF NEOSPORA-CANINUM [J].
ELLIS, J ;
LUTON, K ;
BAVERSTOCK, PR ;
BRINDLEY, PJ ;
NIMMO, KA ;
JOHNSON, AM .
MOLECULAR AND BIOCHEMICAL PARASITOLOGY, 1994, 64 (02) :303-311
[14]  
HOLMDAHL OJM, 1994, FEMS MICROBIOL LETT, V119, P187, DOI 10.1111/j.1574-6968.1994.tb06887.x
[15]  
KAUFMANN H, 1995, SCHWEIZ MED WOCHENSC, V125, P930
[16]   NEOSPORA-CANINUM (PROTOZOA, APICOMPLEXA) INFECTIONS IN MICE [J].
LINDSAY, DS ;
DUBEY, JP .
JOURNAL OF PARASITOLOGY, 1989, 75 (05) :772-779
[17]   INFECTIONS IN MICE WITH TACHYZOITES AND BRADYZOITES OF NEOSPORA-CANINUM (PROTOZOA, APICOMPLEXA) [J].
LINDSAY, DS ;
DUBEY, JP .
JOURNAL OF PARASITOLOGY, 1990, 76 (03) :410-413
[18]   POLYMERASE CHAIN-REACTION FOR DETECTION OF TOXOPLASMA-GONDII [J].
SAVVA, D ;
MORRIS, JC ;
JOHNSON, JD ;
HOLLIMAN, RE .
JOURNAL OF MEDICAL MICROBIOLOGY, 1990, 32 (01) :25-31
[19]  
SAVVA D, 1989, MICROBIOS, V58, P165
[20]   IDENTIFICATION OF THE 4 HUMAN MALARIA PARASITE SPECIES IN FIELD SAMPLES BY THE POLYMERASE CHAIN-REACTION AND DETECTION OF A HIGH PREVALENCE OF MIXED INFECTIONS [J].
SNOUNOU, G ;
VIRIYAKOSOL, S ;
JARRA, W ;
THAITHONG, S ;
BROWN, KN .
MOLECULAR AND BIOCHEMICAL PARASITOLOGY, 1993, 58 (02) :283-292