Detection of Cryptosporidium parvum DNA in formed human feces by a sensitive PCR-based assay including uracil-N-glycosylase inactivation.

Journal of Clinical Microbiology
P GobetA Bonnin

Abstract

We developed a PCR-based method that can be used to identify Cryptosporidium parvum in human feces. Fecal oocysts were concentrated by centrifugation on a sodium chloride gradient and filtration on a nitrocellulose filter prior to DNA extraction and PCR amplification of a 452-bp C. parvum-specific DNA sequence with a protocol including dUTP and uracil-N-glycosylase. All samples obtained from naturally infected humans (n = 10), calves (n = 4), and goats (n = 2) were positive. A 100% detection rate was achieved with both formed and solid stools (n = 10) seeded with 1,000 C. parvum oocysts per g. Procedures based on stool concentration by a modified Ritchie method and subsequent oocyst identification by immunofluorescent labeling or acid-fast staining require concentrations of 50,000 to 500,000 oocysts per g to achieve a 100% detection rate with formed stools. The described PCR-based assay thus has a 50- to 500-fold increase in sensitivity compared to those of the methods commonly used to analyze formed feces.

References

Oct 1, 1992·The American Journal of Tropical Medicine and Hygiene·M A LaxerR J Patel
Dec 1, 1991·The American Journal of Tropical Medicine and Hygiene·M A LaxerR J Patel
Jan 1, 1990·Annals of Internal Medicine·E N JanoffK A Penley
Feb 1, 1990·Epidemiology and Infection·D P Casemore
Jan 1, 1988·Critical Reviews in Microbiology·F G Crawford, S H Vermund
Feb 1, 1988·The Journal of Infectious Diseases·R Soave, W D Johnson
Feb 1, 1986·The Journal of Infectious Diseases·H P Holley, C Dover
Mar 1, 1994·Clinical Infectious Diseases : an Official Publication of the Infectious Diseases Society of America·F BissuelC Trepo
Aug 1, 1994·The Journal of Infectious Diseases·A C WhiteR W Goodgame
Jun 1, 1993·Journal of Clinical Microbiology·J E Rosenblatt, L M Sloan
Feb 1, 1993·Clinical Infectious Diseases : an Official Publication of the Infectious Diseases Society of America·C J FichtenbaumW G Powderly
Jan 1, 1996·The Journal of Infectious Diseases·C L ChappellH L DuPont
Feb 1, 1996·Applied and Environmental Microbiology·X LengR D Oberst
Jul 1, 1996·Journal of Clinical Microbiology·A B BalatbatJ Silva
Apr 1, 1996·Molecular and Biochemical Parasitology·U M MorganR C Thompson

❮ Previous
Next ❯

Citations

Oct 6, 2001·Clinical Infectious Diseases : an Official Publication of the Infectious Diseases Society of America·M KamiH Hirai
Sep 3, 2009·The Korean Journal of Parasitology·Jae-Ran YuWoo-Yoon Park
Sep 13, 2003·Journal of Veterinary Diagnostic Investigation : Official Publication of the American Association of Veterinary Laboratory Diagnosticians, Inc·C K Nielsen, L A Ward
Apr 18, 1998·Applied and Environmental Microbiology·G Pereira M dasR B Lefebvre
Feb 11, 2000·Veterinary Parasitology·T Sréter, I Varga
Oct 9, 1999·Clinical Microbiology Reviews·D P Clark
Mar 13, 1999·Journal of Clinical Microbiology·J LoefflerH Einsele

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.