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Comparison of current methods used to detect Cryptosporidium oocysts in stools
Authors:Shahira A. Ahmed  Panagiotis Karanis
Affiliation:1. Department of Parasitology, Faculty of Medicine, Suez Canal University, Ismailia 41522, Egypt;2. State Key Laboratory of Plateau Ecology and Agriculture, Center for Biomedicine and Infectious Diseases, Qinghai University, Xining, Qinghai 810016, PR China;3. Medical School, University of Cologne, Cologne, Germany
Abstract:In this review all of the methods that are currently in use for the investigation of Cryptosporidium in stool material are highlighted and critically discussed. It appears that more qualifications and background knowledge in this field regarding the diagnosis of the Cryptosporidium parasite is required. Furthermore, there is no standardization for the protocols that are commonly used to either detect oocysts in faeces or to diagnose the Cryptosporidium infection. It is therefore necessary to initiate further education and research that will assist in improving the accuracy of the diagnosis of Cryptosporidium oocysts in the faecal micro-cosmos. Where ambient concentrations of oocysts are low in stool material, detection becomes a formidable task. Procedures for ring tests and the standardization of multi-laboratory testing are recommended. It is also necessary to enhance the routine surveillance capacity of cryptosporidiosis and to improve the safety against it, considering the fact that this disease is under diagnosed and under reported. This review is intended to stimulate research that could lead to future improvements and further developments in monitoring the diagnostic methodologies that will assist in harmonizing Cryptosporidium oocysts in stool diagnosis.
Keywords:ACMV  analine carbol methyl violet  AF  acid flocculation  AOAR  acridine orange, auramine-rhodamine  COWP  CsCl  cesium chloride  ddPCR  droplet digital PCR  DFA  direct immunofluorescence  DGGE  denaturing gradient gel electrophoresis  DMSO-MAF  dimethyl sulfoxide-modified acid-fast  DNA  deoxyribonucleic acid  DSIP  discontinuous sucrose and isopycnic percoll gradients  EIA  enzyme immunoassay  ELISA  enzyme linked immunosorbent assay  EM  electron microscope  EMS  electrophoretic mutation scanning  FA  fluorescent antibody  FC  flow cytometry  FCCS  flow cytometry coupled with cell sorting  FEA  formalin-ethyl acetate  FG  ficoll gradient  FISH  fluorescence in situ hybridization  GBC  glass bead column  gp60  60- kilodalton glycoprotein  HDA  heteroduplex analysis  hsp  heat shock protein  hsp70  70-kilodalton heat shock protein  ICT  immunochromatographic dipstick assay  IFA  indirect immunofluorescence  IMS  immunomagnetic separation  ITS  internal transcribed spacer  KAF  kinyoun acid-fast  K-dichromate  potassium dichromate  LAMP  loop-mediated isothermal amplification  LM  light microscope  LSCM  laser scanning confocal microscope  MAF  modified acid-fast  magnesium sulphate  Opg  oocysts per gram stool  PAS  periodic acid-Schiff  PBDG  potassium bromide discontinuous gradient  PCM  phase contrast microscope  PCR  polymerase chain reaction  PVA  poly-venyle alcohol  qPCR  real-time or quantitative PCR  RFLP  restriction fragment length polymorphism  RIHS  regressive iron hematoxylin-stain  RNA  ribonucleic acid  SAF  acid-formalin fixatives  SEM  scanning electron microscope  SGS  simple gravity sedimentation  SMA  silver methenamine acridine  SSCP  single-strand conformation polymorphism  SSF  sheather sucrose flotation  SSSF  saturated sodium chloride salt flotation  SSU  small-subunit  TEM  transmission electron microscope  ZNAF  Ziehl-Neelsen acid-fast  ZSF  zinc sulphate flotation  Preservation  Detection  Microscopy  Antigenic  Molecular
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