Detection and differentiation of Cryptosporidium by real-time polymerase chain reaction in stool samples from patients in Rio de Janeiro, Brazil

被引:18
作者
Ribeiro Rolando, Roberta Flavia [2 ]
da Silva, Sidnei [3 ]
Saramago Peralta, Regina Helena [1 ]
da Silva, Alexandre Januario [4 ]
Cunha, Flavia de Souza [1 ]
Bello, Alexandre Ribeiro [2 ]
Peralta, Jose Mauro [5 ]
机构
[1] Univ Fed Fluminense, Fac Med, Dept Patol, Niteroi, RJ, Brazil
[2] Univ Estado Rio de Janeiro, Dept Microbiol Imunol & Parasitol, Rio De Janeiro, RJ, Brazil
[3] Fiocruz MS, Inst Pesquisa Clin Evandro Chagas, Rio De Janeiro, RJ, Brazil
[4] Ctr Dis Control & Prevent, Div Parasit Dis & Malaria, Ctr Global Hlth, Atlanta, GA USA
[5] Univ Fed Rio de Janeiro, Inst Microbiol Paulo de Goes, BR-21941 Rio De Janeiro, RJ, Brazil
来源
MEMORIAS DO INSTITUTO OSWALDO CRUZ | 2012年 / 107卷 / 04期
关键词
Cryptosporidium; RT-PCR; molecular epidemiology; MOLECULAR CHARACTERIZATION; SAO-PAULO; PCR ASSAY; PARVUM; CHILDREN; HOMINIS; INFECTIONS; PREVALENCE; PARASITES; ANIMALS;
D O I
10.1590/S0074-02762012000400006
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
This study reports the first genetic characterisation of Cryptosporidium isolates in Brazil using real-time polymerase chain reaction (RT-PCR). A total of 1,197 faecal specimens from children and 10 specimens from human immunodeficiency virus-infected patients were collected between 1999-2010 and screened using microscopy. Forty-eight Cryptosporidium oocyst-positive isolates were identified and analysed using a generic TaqMan assay targeting the 18S rRNA to detect Cryptosporidium species and two other TaqMan assays to identify Cryptosporidium hominis and Cryptosporidium parvum. The 18S rRNA assay detected Cryptosporidium species in all 48 of the stool specimens. The C. parvum TaqMan assay correctly identified five/48 stool samples, while 37/48 stool specimens were correctly amplified in the C. hominis TaqMan assay. The results obtained in this study support previous findings showing that C. hominis infections are more prevalent than C. parvum infections in Brazil and they demonstrate that the TaqMan RT-PCR procedure is a simple, fast and valuable tool for the detection and differentiation of Cryptosporidium species.
引用
收藏
页码:476 / 479
页数:4
相关论文
共 34 条
[1]   Development and evaluation of a real-time PCR assay for quantification of Giardia and Cryptosporidium in sewage samples [J].
Alonso, Jose L. ;
Amoros, Inmaculada ;
Canigral, Irene .
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 2011, 89 (04) :1203-1211
[2]   Heavy cryptosporidial infections in children in northeast Brazil:: comparison of Cryptosporidium hominis and Cryptosporidium parvum [J].
Bushen, Oluma Y. ;
Kohli, Anita ;
Pinkerton, Relana C. ;
Dupnik, Kate ;
Newman, Robert D. ;
Sears, Cynthia L. ;
Fayer, Ronald ;
Lima, Aldo A. M. ;
Guerrant, Richard L. .
TRANSACTIONS OF THE ROYAL SOCIETY OF TROPICAL MEDICINE AND HYGIENE, 2007, 101 (04) :378-384
[3]   Differences in clinical manifestations among Cryptosporidium species and subtypes in HIV-infected persons [J].
Cama, Vitaliano A. ;
Ross, Jennifer M. ;
Crawford, Sara ;
Kawai, Vivian ;
Chavez-Valdez, Raul ;
Vargas, Daniel ;
Vivar, Aldo ;
Ticona, Eduardo ;
Navincopa, Marco ;
Williamson, John ;
Ortega, Ynes ;
Gilman, Robert H. ;
Bern, Caryn ;
Xiao, Lihua .
JOURNAL OF INFECTIOUS DISEASES, 2007, 196 (05) :684-691
[4]   Minireview: Clinical cryptosporidiosis [J].
Chalmers, Rachel M. ;
Davies, Angharad P. .
EXPERIMENTAL PARASITOLOGY, 2010, 124 (01) :138-146
[5]   MOLECULAR CHARACTERIZATION OF Cryptosporidium spp. FROM HIV INFECTED PATIENTS FROM AN URBAN AREA OF BRAZIL [J].
de Lucca, Patricia ;
De Gaspari, Elizabeth Natal ;
Bozzoli, Ligia M. ;
Funada, Mikaela Renata ;
de Souza Silva, Sheila Oliveira ;
Iuliano, Wilma ;
Soares, Rodrigo Martins .
REVISTA DO INSTITUTO DE MEDICINA TROPICAL DE SAO PAULO, 2009, 51 (06) :341-343
[6]   Taxonomy and species delimitation in Cryptosporidium [J].
Fayer, Ronald .
EXPERIMENTAL PARASITOLOGY, 2010, 124 (01) :90-97
[7]   Development of a TaqMan quantitative PCR assay specific for Cryptosporidium parvum [J].
Fontaine, M ;
Guillot, E .
FEMS MICROBIOLOGY LETTERS, 2002, 214 (01) :13-17
[8]   Unique Cryptospoddium population in HIV-infected persons, Jamaica [J].
Gatei, Wangeci ;
Barrett, Donnett ;
Lindo, John F. ;
Eldemire-Shearer, Denise ;
Cama, Vitalliano ;
Xiao, Lihua .
EMERGING INFECTIOUS DISEASES, 2008, 14 (05) :841-843
[9]   Multilocus sequence typing and genetic structure of Cryptosporidium hominis from children in Kolkata, India [J].
Gatei, Wangeci ;
Das, Pradeep ;
Dutta, Phalguni ;
Sen, Abhik ;
Cama, Vitaliano ;
Lal, Altaf A. ;
Xiao, Lihua .
INFECTION GENETICS AND EVOLUTION, 2007, 7 (02) :197-205
[10]   Cryptosporidiosis: Prevalence, genotype analysis, and symptoms associated with infections in children in Kenya [J].
Gatei, Wangeci ;
Wamae, Claire N. ;
Mbae, Cecilia ;
Waruru, Anthony ;
Mulinge, Erastus ;
Waithera, Tabitha ;
Gatika, Simon M. ;
Kamwati, Stanely K. ;
Revathi, Gunturu ;
Hart, Charles A. .
AMERICAN JOURNAL OF TROPICAL MEDICINE AND HYGIENE, 2006, 75 (01) :78-82