Hi‐PCR® Protozoan Parasite Multiplex Probe PCR Kit

Description:

Haemoparasitic infections pose serious problems for the health and management of domestic cattle in tropical and subtropical regions of the world. Among these diseases, babesiosis, tropical theileriosis, anaplasmosis and trypanosomosis caused by Babesia bigemina, Theileria annulata, Anaplasma marginale and Trypanosoma evansi respectively, are the most economically important vector borne diseases encountered by cattle. These diseases are responsible for high level of morbidity and mortality in susceptible population and thereby causing huge economic losses to the dairy industry. Clinical signs associated with above diseases are overlapping and may include pyrexia, anorexia, anemia, icterus, hemoglobinuria, weight loss, reduced milk production. Co‐infection of these parasites in animals is a common feature in most parts of the world. Hence, timely and specific diagnosis of the causative agent before implementation of appropriate treatment and related control measures is very important. Polymerase Chain Reaction (PCR) technology will help to provide a quick, accurate and highly specific diagnosis of economically important diseases of livestock. Among the different PCR protocols for pathogen detection, multiplex PCR assay offers an advantage of detecting multiple pathogens in a single reaction and has been employed to detect mixed infections of haemoparasites.
NOTE: Hi‐PCR Protozoan Parasite Multiplex Probe PCR Kit is for in vitro use only.

Intended Use:

Hi‐PCR® Protozoan Parasite Multiplex Probe PCR Kit is intended for use by qualified laboratory personnel trained in Real‐Time PCR. The kit is recommended for sensitive and specific detection of Anaplasma, Babesia, Theileria and Trypanosoma in a single tube assay.

Principle:

Real‐time polymerase chain reaction, also called quantitative Polymerase Chain Reaction (qPCR) or kinetic Polymerase Chain Reaction, is a laboratory technique based on the principle of PCR. This technique is used to amplify a targeted DNA sequence by use of hydrolysis probes that are short oligonucleotides that have a fluorescent reporter dye attached to the 5' end and a quencher dye to the 3' end. Hi‐PCR® Protozoan Parasite Multiplex Probe PCR Kit is designed to detect Theileria, Anaplasma, Trypanosoma and Babesia in FAM, Texas Red, Cy5 and Cy5.5 channel, respectively, and the Internal Control in JOE channel in a single tube reaction. The kit allows sensitive and specific detection of all targeted Protozoan Parasites.

Negative Control

A negative control is needed to ensure that the reagents, equipment, and environment used in the assay is not contaminated. In this reaction, nuclease free water is used as the template.

Positive control

This is a control reaction using a known template (target pathogen). A positive control is usually used to ensure proper and intended functioning of all the reagents and is recommended to be used in every run to assess optimal performance.

Internal Control

This is a control sequence which is amplified in the same reaction tube along with the target sequence but detected with a different primer (i.e. Multiplex PCR). An internal control is often used to detect the failure of amplification in cases where the target sequence is not amplified.