PCRmax Cyclospora cayetanensis Instruction Manual

Heat shock protein 70 (HSP70) gene
Cyclospora cayetanensis
PCRmax Ltd qPCR test
150 tests
For general laboratory and research use only
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Cyclospora cayetanensis is an intestinal apicomplexan, cyst-forming, protozoan parasite. It causes chronic diarrhoeal disease in humans called cyclosporiasis, commonly referred to as traveller’s diarrhoea. It was first reported in 1979 but was officially classified as C. cayetanensis in the early 1990s due to increased world wide incidence of the disease. C. cayetanensis is coccidian with spherical oocysts of 7.5-10µM in diameter and a bilayered cell wall with an outer threadlike coat. On sporulation, the oocyst has two sporocysts that are approximately 4 by 6.3µm and each sporocyst contains two folded sporozoites of approximately 1.2 by 9µm.
The primary mode of transmission is the faecal-oral route and the parasite spreads indirectly via the environment through oocyst contamination of water, food or soil. Upon ingestion the host gastrointestinal environment causes the oocysts to excyst and release sporozoites which invade the epithelial cells of the small intestine, particularly the jejunum. Some of the parasitic cells undergo a form of asexual reproduction, merogony, which results in many daughter merozoites. The merozoites go on to infect further host cells and induce further rounds of merogony. Alternatively, a sexual reproductive stage may occur to produce further oocysts that are shed in the host faecal matter and act to spread the infection further. After one to two weeks, the shed oocysts become infectious and the pathogen can infect a new host. Due to this delay, direct person-to-person spread of C. cayetanensis is unlikely. Environmental factors such as climate, temperature and moisture determine when the oocytes become infectious.
Symptoms begin approximately one week after ingestion of contaminated food or water. They include frequent diarrhoea, weight loss, flatulence, fatigue, abdominal cramping, anorexia, vomiting and nausea. Infection can be asymptomatic and the disease is usually self-limiting. Although the disease is not fatal, patients can suffer from relapse. In severe cases, symptoms can last more than a month if left untreated and the disease is commonly observed in AIDS patients.
Introduction to Cyclospora cayetanensis
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MIN
MAX
The PCR Max qPCR Kit for Cyclospora cayetanensis (C.cayetanensis) genomes is designed for the in vitro quantification of C.cayetanensis genomes. The kit is designed to have the broadest detection profile possible whilst remaining specific to the C. cayetanensis genome.
The primers and probe sequences in this kit have 100% homology with a broad range of C.cayetanensis sequences based on a comprehensive bioinformatics analysis.
If you require further information, or have a specific question about the detection profile of this kit then please send an e.mail to help@pcrmax.com and our bioinformatics team will answer your question.
Specificity
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Kit Contents
• C.cayetanensis specific primer/probe mix (150 reactions BROWN)
FAM labelled
• C.cayetanensis positive control template (for Standard curve RED)
• Internal extraction control primer/probe mix (150 reactions BROWN)
VIC labelled as standard
• Internal extraction control DNA (150 reactions BLUE)
• Endogenous control primer/probe mix (150 reactions BROWN)
FAM labelled
• RNAse/DNAse free water (WHITE)
for resuspension of primer/probe mixes and internal extraction control DNA
• Template preparation buffer (YELLOW)
for resuspension of positive control template and standard curve preparation
Reagents and equipment to be supplied by the user
Real-Time PCR Instrument
DNA extraction kit
This kit designed to work well with all processes that yield high quality DNA with minimal PCR inhibitors.
Lyophilised 2x qPCR Mastermix
This kit is designed to be compatible with all commercially available Mastermixes that run with standard cycling conditions.
Pipettors and Tips
Vortex and centrifuge
Thin walled 1.5 ml PCR reaction tubes
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