PCR-coupled sequencing achieves specific diagnosis of onchocerciasis in a challenging clinical case, to underpin effective treatment and clinical management

Crowe, Amy; Koehler, Anson V.; Sheorey, Harsha; Tolpinrud, Anita; Gasser, Robin B.

doi:10.1016/j.meegid.2018.09.012

Cited by 4 publications

(6 citation statements)

References 9 publications

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“…DNA-based detection methods are more sensitive methods to detect Onchocerca spp. [38][39][40] ; however, the efficiency of these methods (specificity and sensitivity) depends on the quality of the preserved specimen and DNA template, set of oligonucleotides selected, and the size of fragments amplified. For instance, the O-150 locus, the standard target used for the detection of O. volvulus in programs for monitoring and elimination of onchocerciasis, has intraspecies variation that can affect the sensitivity of PCR methods.…”

Section: Discussionmentioning

confidence: 99%

Duplex Real-Time PCR Assay for Clinical Differentiation of Onchocerca lupi and Onchocerca volvulus

Almeida

Nascimento

Mathison

et al. 2020

The American Journal of Tropical Medicine and Hygiene

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In the United States and Europe, human onchocerciasis is a rare disease caused by zoonotic or anthropophilic parasites in the genus Onchocerca. The zoonotic species identified in focal areas of Europe and United States is Onchocerca lupi, and Onchocerca volvulus, the anthroponotic species, may be found among people who had lived in endemic areas of Africa, the Arabian Peninsula, or Latin America. Onchocerciasis due to O. lupi is an emergent parasitic disease, with limited diagnostic methods, in addition to the lack of information on its biology, transmission, and epidemiology. Cutaneous nodules are the disease's most prevalent manifestation but lack diagnostic specificity. To address the diagnosis of onchocerciasis at reference laboratories, we developed a duplex TaqMan real-time PCR (qPCR) method, targeting the cytochrome oxidase subunit I locus which has species-specific probes to identify and differentiate O. lupi from O. volvulus. We determined the performance of the duplex with a panel of 45 samples: 11 positives for O. lupi, six for O. volvulus, five samples with negative results for Onchocerca spp., and 23 non-Onchocerca nematodes. The duplex qPCR correctly detected 10 of 11 O. lupi-and six of six O. volvulus-positive specimens. The new duplex assay allowed the simultaneous detection and discrimination of O. lupi and O. volvulus in clinical specimens, expediting and facilitating the clinical diagnosis of O. lupi in non-endemic settings where the disease is an infrequent finding.

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Section: Discussionmentioning

confidence: 99%

Duplex Real-Time PCR Assay for Clinical Differentiation of Onchocerca lupi and Onchocerca volvulus

Almeida

Nascimento

Mathison

et al. 2020

The American Journal of Tropical Medicine and Hygiene

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“…After the worm was extracted, experts were convinced that the morphology was most consistent with Onchocerca species. A PCR-based sequencing method confirmed it to be O. volvulus (1).…”

Section: Case 2: Lump In the Axilla (Brisbane Qld Australia)mentioning

confidence: 93%

“…The author acts as a coordinator of the group. The team has helped diagnose a few unusual parasites, leading to publications [1][2][3][4]. Over the past decade or two, the teaching of laboratory diagnostic parasitology has been neglected in Australasia because parasitic infections are relatively uncommon.…”

Section: Who Is the Team?mentioning

confidence: 99%

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e-Diagnosis in Medical Parasitology

Sheorey

2020

TropicalMed

Self Cite

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Over the past decade or two, the teaching of laboratory diagnostic parasitology has been neglected in Australasia, as parasitic infections are relatively uncommon. As a consequence, expertise in medical parasitology is dwindling. A team of international experts (including Professor John Goldsmid) has been formed to help in the diagnosis of human parasitic infections. The team includes experts from Australia, Europe, South Africa and the USA. Some senior members of the team are excellent morphologists, and we have both human and veterinary parasitologists who help with molecular diagnosis in difficult cases.

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“…Two of the more common tests are the OV-16 antibody test (38) and the OV luciferase immunoprecipitation system (39). There are no specific molecular assays that target O. volvulus for routine clinical diagnosis; however, PCR amplification and sequencing of the cox-1 gene has been used in clinical care in Australia to diagnosis onchocerciasis in specimens containing a worm visualized by microscopy (40).…”

Section: Species-specific Informationmentioning

confidence: 99%

Diagnostic Identification and Differentiation of Microfilariae

2019

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The morphologic similarities of the microfilariae and their infrequency in clinical specimens in settings of endemicity present challenges to clinical laboratories in maintaining competence for accurate identification and differentiation. We present here a review of the primary filarial nematodes causing human infection, including an illustrated key, which we hope will improve the diagnostic capabilities of hematologists, microbiologists, medical technologists, and similarly qualified laboratorians.

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PCR-coupled sequencing achieves specific diagnosis of onchocerciasis in a challenging clinical case, to underpin effective treatment and clinical management

Cited by 4 publications

References 9 publications

Duplex Real-Time PCR Assay for Clinical Differentiation of Onchocerca lupi and Onchocerca volvulus

Duplex Real-Time PCR Assay for Clinical Differentiation of Onchocerca lupi and Onchocerca volvulus

e-Diagnosis in Medical Parasitology

Diagnostic Identification and Differentiation of Microfilariae

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