Detection and differentiation of Cryptosporidium by real-time polymerase chain reaction in stool samples from patients in Rio de Janeiro, Brazil.
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.
Departamento de Microbiologia, Imunologia e Parasitologia, Universidade do Estado do Rio de Janeiro, Rio de Janeiro, RJ, Brazil.
Roberta Flávia Ribeiro
RF
Rolando
Sidnei da
Sd
Silva
Regina Helena Saramago
RH
Peralta
Alexandre Januário da
AJ
Silva
Flavia de Souza
Fde S
Cunha
Alexandre Ribeiro
AR
Bello
José Mauro
JM
Peralta
D017088:AIDS-Related Opportunistic Infections / Q000175:diagnosis / Q000453:epidemiology / Q000469:parasitology*; D001938:Brazil / Q000453:epidemiology; D002648:Child; D003457:Cryptosporidiosis / Q000175:diagnosis / Q000453:epidemiology / Q000469:parasitology*; D003458:Cryptosporidium / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D016785:Cryptosporidium parvum / Q000145:classification / Q000235:genetics / Q000302:isolation & purification; D016054:DNA, Protozoan / Q000032:analysis; D004275:DNA, Ribosomal / Q000032:analysis; D005243:Feces / Q000469:parasitology*; D006801:Humans; D015995:Prevalence; D012337:RNA, Ribosomal, 18S / Q000032:analysis; D060888:Real-Time Polymerase Chain Reaction; D012680:Sensitivity and Specificity
D016054:DNA, Protozoan; D004275:DNA, Ribosomal; D012337:RNA, Ribosomal, 18S