The pathogenic Leptospira qPCR detection kit is based on the fast, sensitive, and proven primers/probe qPCR technique. The kit contains primers and a probe for detecting a highly specific sequence present on the DNA of pathogenic Leptospira. This kit of Sylphium molecular ecology is designed to have a sensitivity of only one molecule per reaction in environmental samples.
Disease information
Leptospirosis is a blood infection caused by corkscrew-shaped bacteria called Leptospira. Signs and symptoms can range from none to mild such as headaches, muscle pains, and fevers to severe with bleeding from the lungs or meningitis.In a more severe form, known as Weil’s disease, it causes the person to turn yellow, have kidney failure, and bleed.If it also causes bleeding into the lungs then it is known as severe pulmonary haemorrhage syndrome.
More than ten different genetic types of Leptospira may cause disease in humans.Both wild and domestic animals can spread the disease, with the most common being rodents. The bacteria are spread through animal urine, or water and soil contaminated with animal urine coming into contact with breaks in the skin, eyes, mouth, or nose.In the developing countries the disease occurs most commonly in farmers and low-income people who live in areas with poor sanitation. In developed countries, it occurs during heavy downpour and in those involved in outdoor activities in warm and wet areas. Diagnosis is typically by testing for antibodies against the bacteria or finding bacterial DNA.
In English: Weil’s disease
In German: Weil-Krankheit
In French: Maladie de Weil
In Dutch: Ziekte van Weil
Primer design
Primers and probe and the qPCR buffer were designed to have:
- Highest possible sensitivity (1 DNA copy per reaction). Environmental water samples contain normally very low amounts of target DNA.
- Strong fluorescence signal with low background noise. Isolated environmental samples contain residues of naturally occurring auto fluoresce substances that will interfere with the measurements. A strong fluorescence signal from the analyses is required for these kind of samples.
- 100% specificity. Isolated DNA from environmental samples contains billions of DNA fragments from bacteria, protozoa, plants, animals and must be taken in account during primer/probe design. This is validated by experimental and bioinformatical studies.
The kit is developed and optimized to be used on eDNA isolates purified using the eDNA isolation kit (#SYL002) from Sylphium molecular ecology.
Kit contents
The kit contains materials for an 8-fold analyses on 20 samples including all necessary controls.
- Positive control (cloned pathogenic Leptospira DNA)
- 2x Sylphium qPCR mix (100 reactions) without primers and probes
- 2x Primer/probe mix (100 reactions) for detection of pathogenic Leptospira (FAM dye)
- 1x Taq DNA polymerase (200 reactions)
- 2 ml dilution solution (S6)
- Batch validation document
Manual