Diagnosis of protozoa diarrhoea in Campylobacter patients increases markedly with molecular techniques.

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dc.citation.issue5
dc.citation.volume3
dc.contributor.authorHayman DTS
dc.contributor.authorGarcia-Ramirez JC
dc.contributor.authorPita A
dc.contributor.authorVelathanthiri N
dc.contributor.authorKnox MA
dc.contributor.authorOgbuigwe P
dc.contributor.authorBaker MG
dc.contributor.authorRostami K
dc.contributor.authorDeroles-Main J
dc.contributor.authorGilpin BJ
dc.contributor.editorStandley C
dc.coverage.spatialUnited States
dc.date.accessioned2024-07-23T21:49:51Z
dc.date.available2024-07-23T21:49:51Z
dc.date.issued2023-05-30
dc.description.abstractCryptosporidium and Giardia are major causes of diarrhoea globally, and two of the most notified infectious diseases in New Zealand. Diagnosis requires laboratory confirmation carried out mostly via antigen or microscopy-based techniques. However, these methods are increasingly being superseded by molecular techniques. Here we investigate the level of protozoa detection by molecular methods in campylobacteriosis cases missed through antigen-based assays and investigate different molecular testing protocols. We report findings from two observational studies; the first among 111 people during a Campylobacter outbreak and the second during normal surveillance activities among 158 people presenting with diarrhoea and a positive Campylobacter test, but negative Cryptosporidium and Giardia antigen-based test results. The molecular methods used for comparison were in-house end-point PCR tests targeting the gp60 gene for Cryptosporidium and gdh gene for Giardia. DNA extraction was performed with and without bead-beating and comparisons with commercial real-time quantitative (qPCR) were made using clinical Cryptosporidium positive sample dilutions down to 10-5. The Cryptosporidium prevalence was 9% (95% CI: 3-15; 10/111) and Giardia prevalence 21% (95% CI: 12-29; 23/111) in the 111 Campylobacter outbreak patients. The Cryptosporidium prevalence was 40% (95% CI: 32-48; 62/158) and Giardia prevalence 1.3% (95% CI: 0.2-4.5; 2/158) in the 158 routine surveillance samples. Sequencing identified Cryptosporidium hominis, C. parvum, and Giardia intestinalis assemblages A and B. We found no statistical difference in positive test results between samples using end-point PCR with or without bead-beating prior to DNA extraction, or between the in-house end-point PCR and qPCR. The qPCR Ct value was 36 (95% CI: 35-37) for 1 oocyst, suggesting a high limit of detection. In conclusion in surveillance and outbreak situations we found diagnostic serology testing underdiagnoses Cryptosporidium and Giardia coinfections in Campylobacter patients, suggesting the impact of protozoa infections may be underestimated through underdiagnosis using antigen-based assays.
dc.description.confidentialfalse
dc.edition.editione0001527
dc.format.paginatione0001527-
dc.identifier.author-urlhttps://www.ncbi.nlm.nih.gov/pubmed/37252910
dc.identifier.citationHayman DTS, Garcia-Ramirez JC, Pita A, Velathanthiri N, Knox MA, Ogbuigwe P, Baker MG, Rostami K, Deroles-Main J, Gilpin BJ. (2023). Diagnosis of protozoa diarrhoea in Campylobacter patients increases markedly with molecular techniques.. PLOS Glob Public Health. 3. 5. (pp. e0001527-).
dc.identifier.doi10.1371/journal.pgph.0001527
dc.identifier.eissn2767-3375
dc.identifier.elements-typejournal-article
dc.identifier.issn2767-3375
dc.identifier.piiPGPH-D-22-02068
dc.identifier.urihttps://mro.massey.ac.nz/handle/10179/70289
dc.languageeng
dc.publisherPublic Library of Science (PLoS)
dc.publisher.urihttps://www.sciencedirect.com/science/article/pii/S2352771423001374?
dc.relation.isPartOfPLOS Glob Public Health
dc.rights(c) 2023 The Author/s
dc.rightsCC BY 4.0
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/
dc.titleDiagnosis of protozoa diarrhoea in Campylobacter patients increases markedly with molecular techniques.
dc.typeJournal article
pubs.elements-id461865
pubs.organisational-groupOther
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