Optimization of a rapid nonisotopic DNA probe assay for Plasmodium falciparum in the Gambia

McLaughlin, Gerald L.; Decrind, C.; Dayal-Drager, Renu; Hassan-King, Musa; Subramanian, Sathish; Greenwood, Brian

doi:10.1128/jcm.29.7.1517-1519.1991

Cited by 11 publications

(2 citation statements)

References 17 publications

(21 reference statements)

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“…The probes were highly specific but did not detect low levels of parasitemia (Ͻ500 parasites per l). When the synthetic probe conjugated to alkaline phosphatase was used and a protocol for microtiter plate-based blood lysis with detergent and protease followed by filtration was optimized for blood processing and storage and filter treatment (73,75), the sensitivity of the assay increased from 84 to 95%, while specificity remained excellent compared with microscopy. The patients were representatives of populations with high levels of parasitemia (The Gambia) and low levels of parasitemia with other species of Plasmodium present (Madagascar).…”

Section: P Falciparummentioning

confidence: 99%

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DNA probes and PCR for diagnosis of parasitic infections

Weiss

1995

Clin Microbiol Rev

130

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DNA probe and PCR-based assays to identify and detect parasites are technically complex; however, they have high sensitivity, directly detect parasites independent of the immunocompetence or previous clinical history of the patient, and can distinguish between organisms that are morphologically similar. Diagnosis of parasites is often based on direct detection by microscopy, which is insensitive and laborious and can lack specificity. Most PCR-based assays were more sensitive than DNA probe assays. The development of PCR-based diagnostic assays requires multiple steps following the initial selection of oligonucleotide primers and reporter probe. Generally, the ability to detect the DNA of one parasite was attained by PCR; however, advances in the preparation of samples for PCR (extraction of DNA while removing PCR inhibitors) will be required to achieve that sensitivity with human specimens. Preliminary PCR systems have been developed for many different parasites, yet few have been evaluated with a large number of clinical specimens and/or under field conditions. Those evaluations are essential for determination of clinical and field utility and performance and of the most appropriate application of the assay. Several situations in which PCR-based diagnosis will result in epidemiologic, medical, or public health advances have been identified.

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Section: P Falciparummentioning

confidence: 99%

“…Direct spotting onto membranes was the easiest and simplest procedure; however, heme and its breakdown products are known PCR inhibitors, and blood also can contain unknown PCR inhibitors. The lysis and filtration protocol was similar to that successfully used in the field with a DNA probe assay (73,75).…”

Section: P Falciparummentioning

confidence: 99%