Identification of Entamoeba histolytica and E. dispar cysts in stool by polymerase chain reaction.
An attempt to identify cysts of Entamoeba histolytica and E. dispar in human stool was conducted by polymerase chain reaction (PCR) using two sets of primers (p11 plus p12 and p13 plus p14) specific for either species of ameba. The cysts in stool specimens obtained from 12 infected individuals were concentrated, freeze-thawed, and treated with Triton X-100 before their examination by PCR. The results of PCR on the cysts were generally consistent with data obtained by PCR on ameba trophozoites hatched from the cysts, by zymodeme analysis, and by enzyme-linked immunosorbent assay (ELISA) and with clinical findings. This PCR was negative for the stool containing large numbers of cysts of either E. coli, E. hartmanni, or Giardia lamblia as well as for the stool specimens obtained from uninfected individuals. The ameba cyst in stool processed using the present method was effective for the PCR analysis even after 1 month of storage at 4 degrees C. The present PCR was sensitive enough to detect ten cysts of either of the amebae.
Sanuki
J
J
Department of Tropical Medicine and Parasitology, School of Medicine, Keio University, Tokyo, Japan.
Asai
T
T
Okuzawa
E
E
Kobayashi
S
S
Takeuchi
T
T
D000818:Animals; D016054:DNA, Protozoan / Q000235:genetics; D004747:Entamoeba / Q000235:genetics* / Q000254:growth & development / Q000302:isolation & purification; D004748:Entamoeba histolytica / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification; D004749:Entamoebiasis / Q000175:diagnosis*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005243:Feces / Q000469:parasitology*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013045:Species Specificity
D016054:DNA, Protozoan