Direct and sensitive detection of a pathogenic protozoan, Toxoplasma gondii, by polymerase chain reaction.
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TLDR
A combination of sensitivity and specificity should make detection of the B1 gene based on polymerase chain reaction amplification a very useful method for diagnosis of toxoplasmosis both in immunocompromised hosts and in congenitally infected fetuses.Abstract:
We applied the polymerase chain reaction to detection of the pathogenic protozoan Toxoplasma gondii based on our identification of a 35-fold-repetitive gene (the B1 gene) as a target. Using this procedure, we were able to amplify and detect the DNA of a single organism directly from a crude cell lysate. This level of sensitivity also allowed us to detect the B1 gene from purified DNA samples containing as few as 10 parasites in the presence of 100,000 human leukocytes. This is representative of the maximal cellular infiltration (10(5)/ml) in 1 ml of cerebrospinal fluid obtained from patients with toxoplasmic encephalitis. The B1 gene is present and conserved in all six T. gondii strains tested to date, including two isolates from patients with acquired immunodeficiency syndrome. No signal was detected by using this assay and DNAs from a variety of other organisms, including several which might be found in the central nervous system of an immunocompromised host. This combination of sensitivity and specificity should make detection of the B1 gene based on polymerase chain reaction amplification a very useful method for diagnosis of toxoplasmosis both in immunocompromised hosts and in congenitally infected fetuses.read more
Citations
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The polymerase chain reaction. A new method of using molecular genetics for medical diagnosis.
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Epidemiology of and Diagnostic Strategies for Toxoplasmosis
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Toxoplasmic Encephalitis in AIDS
TL;DR: This commentary comprises an update of their initial review of T. gondii and a presentation of the current approaches to diagnosing and managing toxoplasmic encephalitis in HIV-infected patients.
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Toxoplasma gondii: transmission, diagnosis and prevention
TL;DR: Toxoplasmosis is one of the most common parasitic infections of man and other warm-blooded animals and can cause blindness and mental retardation in congenitally infected children and devastating disease in immunocompromised individuals.
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TL;DR: A thermostable DNA polymerase was used in an in vitro DNA amplification procedure, the polymerase chain reaction, which significantly improves the specificity, yield, sensitivity, and length of products that can be amplified.
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TL;DR: Two new methods were used to establish a rapid and highly sensitive prenatal diagnostic test for sickle cell anemia, using primer-mediated enzymatic amplification of specific beta-globin target sequences in genomic DNA, resulting in the exponential increase of target DNA copies.
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