PCRmax Babesia bovis Instruction Manual

18S ribosomal RNA (18S) gene
Babesia bovis
PCRmax Ltd qPCR test
TM
For general laboratory and research use only
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Babesia bovis is a single-celled protozoan parasite of the Apicomplexa phylum measuring
1.5 x 2.4μm in size. The linear DNA genome contains 4 chromosomes and is 8.2Mbp in length. Infection with B. bovis causes a haemolytic anaemia in bovine known as Babesiosis.
B. bovis uses ticks of the Ixodidae family as a vector and infects the vertebrate host via tick feeding. The parasite enters the host in trophozoite ("ring") form and invades the red blood cells by apicoplast action, where it multiplies through binary fission. This increase in parasitic number results in rupture of the erythrocytes causing anaemia and facilitating protozoan spread to other cells. The merozoite form is ingested by uninfected ticks when feeding and settle to the midgut where they multiply. B. bovis then migrate to the salivary glands, ready for injection into a new host.
An incubation period of 1-3 weeks in bovine precedes a high fever which lasts several days and is followed by onset of other symptoms. These include increased respiratory rate, reluctance to move and congestion of the conjunctiva and mucus membranes. As the number of disrupted red blood cells increases anaemia occurs resulting in jaundice. The central nervous system may also be affected causing convulsions, paralysis and coma. This disease is severe with a high fatality rate which increases when the nervous system is involved. Treatment with Imidocarb can increase the survival rates when given early in the infection course but supportive therapies such as blood transfusions may be necessary if treatment is delayed. A live vaccine is available and can provide long lasting immunity.
Human infection is rare and seen mainly among the elderly or those with compromised immune systems, especially splenectomy patients. Infection may cause fever, prostration, jaundice, anaemia and haemoglobulinuria and is usually treated with malaria remedies.
Introduction to Babesia bovis
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MIN
MAX
Our kit for Babesia bovis has been designed for the specific and exclusive in vitro quantification of this species . The 18S ribosomal gene, is the ideal target to achieve a broad based detection profile for all isolates within this species. The primers and probe sequences in this kit have 100% homology with a broad range of clinically relevant reference sequences based on a comprehensive bioinformatics analysis. Other closely related Babesia species are not detected.
The PCR Max qPCR Kit for Babesia bovis (B.bovis ) genomes is designed for the in vitro quantification of B.bovis genomes. The kit is designed to have the broadest detection profile possible whilst remaining specific to the B.bovis genome.
The primers and probe sequences in this kit have 100% homology with a broad range of B.bovis 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
• B.bovis specific primer/probe mix (150 reactions BROWN)
FAM labelled
• B.bovis 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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