Comparison of Three Real-Time PCR Assays Targeting the SSU rRNA Gene, the COWP Gene and the DnaJ-Like Protein Gene for the Diagnosis of <i>Cryptosporidium</i> spp. in Stool Samples
As qualified microscopy of enteric parasitoses as defined by high diagnostic accuracy is difficult to maintain in non-endemic areas due to scarce opportunities for practicing with positive sample materials, molecular diagnostic options provide less investigator-dependent alternatives. Here, we compa...
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| Format: | Article |
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MDPI AG
2021-09-01
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| Series: | Pathogens |
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| Online Access: | https://www.mdpi.com/2076-0817/10/9/1131 |
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| author | Felix Weinreich Andreas Hahn Kirsten Alexandra Eberhardt Torsten Feldt Fred Stephen Sarfo Veronica Di Cristanziano Hagen Frickmann Ulrike Loderstädt |
| author_facet | Felix Weinreich Andreas Hahn Kirsten Alexandra Eberhardt Torsten Feldt Fred Stephen Sarfo Veronica Di Cristanziano Hagen Frickmann Ulrike Loderstädt |
| author_sort | Felix Weinreich |
| collection | DOAJ |
| description | As qualified microscopy of enteric parasitoses as defined by high diagnostic accuracy is difficult to maintain in non-endemic areas due to scarce opportunities for practicing with positive sample materials, molecular diagnostic options provide less investigator-dependent alternatives. Here, we compared three molecular targets for the real-time PCR-based detection of <i>Cryptosporidium</i> spp. From a population of 1000 individuals comprising both Ghanaian HIV (human immunodeficiency virus) patients and military returnees after deployment in the tropics, stool samples were assessed for <i>Cryptosporidium</i> spp. by real-time PCR targeting the small subunit ribosomal RNA (SSU rRNA) gene, the <i>Cryptosporidium</i> oocyst wall (COWP) gene, and the DnaJ-like protein gene (DnaJ), respectively. In declining order, sensitivity of 100% for the SSU rRNA gene PCR, 90.0% for the COWP PCR and 88.8% for the DnaJ PCR, respectively, as well as specificity of 99.6% for the COWP PCR and 96.9% for both the SSU rRNA gene PCR and the DnaJ PCR, respectively, were recorded. Substantial agreement (kappa value 0.663) between the three assays was observed. Further, an accuracy-adjusted <i>Cryptosporidium</i> spp. prevalence of 6.0% was calculated for the study population. In conclusion, none of the assessed real-time PCR assays were associated with perfect test accuracy. However, a combination of highly sensitive SSU rRNA gene PCR for screening purposes and more specific COWP PCR for confirmatory testing should allow reliable diagnosis of <i>Cryptosporidium</i> spp. in stool samples even in low prevalence settings. |
| first_indexed | 2024-03-10T07:19:32Z |
| format | Article |
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| institution | Directory Open Access Journal |
| issn | 2076-0817 |
| language | English |
| last_indexed | 2024-03-10T07:19:32Z |
| publishDate | 2021-09-01 |
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| record_format | Article |
| series | Pathogens |
| spelling | doaj.art-b3f11f2323f74cefb0880197dbaeba612023-11-22T14:42:13ZengMDPI AGPathogens2076-08172021-09-01109113110.3390/pathogens10091131Comparison of Three Real-Time PCR Assays Targeting the SSU rRNA Gene, the COWP Gene and the DnaJ-Like Protein Gene for the Diagnosis of <i>Cryptosporidium</i> spp. in Stool SamplesFelix Weinreich0Andreas Hahn1Kirsten Alexandra Eberhardt2Torsten Feldt3Fred Stephen Sarfo4Veronica Di Cristanziano5Hagen Frickmann6Ulrike Loderstädt7Department of Microbiology and Hospital Hygiene, Bundeswehr Hospital Hamburg, 20359 Hamburg, GermanyDepartment of Medical Microbiology, Virology and Hygiene, University Medicine Rostock, 18057 Rostock, GermanyInstitute of Transfusion Medicine, University Medical Center Hamburg-Eppendorf, 20251 Hamburg, GermanyDepartment of Gastroenterology, Hepatology and Infectious Diseases, University Medical Center Düsseldorf, 40225 Düsseldorf, GermanyDepartment of Medicine, Kwame Nkrumah University of Science and Technology, Kumasi 00233, GhanaInstitute of Virology, Faculty of Medicine and University Hospital of Cologne, University of Cologne, 50935 Cologne, GermanyDepartment of Microbiology and Hospital Hygiene, Bundeswehr Hospital Hamburg, 20359 Hamburg, GermanyDepartment of Hospital Hygiene & Infectious Diseases, University Medicine Göttingen, 37075 Göttingen, GermanyAs qualified microscopy of enteric parasitoses as defined by high diagnostic accuracy is difficult to maintain in non-endemic areas due to scarce opportunities for practicing with positive sample materials, molecular diagnostic options provide less investigator-dependent alternatives. Here, we compared three molecular targets for the real-time PCR-based detection of <i>Cryptosporidium</i> spp. From a population of 1000 individuals comprising both Ghanaian HIV (human immunodeficiency virus) patients and military returnees after deployment in the tropics, stool samples were assessed for <i>Cryptosporidium</i> spp. by real-time PCR targeting the small subunit ribosomal RNA (SSU rRNA) gene, the <i>Cryptosporidium</i> oocyst wall (COWP) gene, and the DnaJ-like protein gene (DnaJ), respectively. In declining order, sensitivity of 100% for the SSU rRNA gene PCR, 90.0% for the COWP PCR and 88.8% for the DnaJ PCR, respectively, as well as specificity of 99.6% for the COWP PCR and 96.9% for both the SSU rRNA gene PCR and the DnaJ PCR, respectively, were recorded. Substantial agreement (kappa value 0.663) between the three assays was observed. Further, an accuracy-adjusted <i>Cryptosporidium</i> spp. prevalence of 6.0% was calculated for the study population. In conclusion, none of the assessed real-time PCR assays were associated with perfect test accuracy. However, a combination of highly sensitive SSU rRNA gene PCR for screening purposes and more specific COWP PCR for confirmatory testing should allow reliable diagnosis of <i>Cryptosporidium</i> spp. in stool samples even in low prevalence settings.https://www.mdpi.com/2076-0817/10/9/1131<i>Cryptosporidium</i> spp.real-time PCRtest comparisonlatent class analysissensitivityspecificity |
| spellingShingle | Felix Weinreich Andreas Hahn Kirsten Alexandra Eberhardt Torsten Feldt Fred Stephen Sarfo Veronica Di Cristanziano Hagen Frickmann Ulrike Loderstädt Comparison of Three Real-Time PCR Assays Targeting the SSU rRNA Gene, the COWP Gene and the DnaJ-Like Protein Gene for the Diagnosis of <i>Cryptosporidium</i> spp. in Stool Samples Pathogens <i>Cryptosporidium</i> spp. real-time PCR test comparison latent class analysis sensitivity specificity |
| title | Comparison of Three Real-Time PCR Assays Targeting the SSU rRNA Gene, the COWP Gene and the DnaJ-Like Protein Gene for the Diagnosis of <i>Cryptosporidium</i> spp. in Stool Samples |
| title_full | Comparison of Three Real-Time PCR Assays Targeting the SSU rRNA Gene, the COWP Gene and the DnaJ-Like Protein Gene for the Diagnosis of <i>Cryptosporidium</i> spp. in Stool Samples |
| title_fullStr | Comparison of Three Real-Time PCR Assays Targeting the SSU rRNA Gene, the COWP Gene and the DnaJ-Like Protein Gene for the Diagnosis of <i>Cryptosporidium</i> spp. in Stool Samples |
| title_full_unstemmed | Comparison of Three Real-Time PCR Assays Targeting the SSU rRNA Gene, the COWP Gene and the DnaJ-Like Protein Gene for the Diagnosis of <i>Cryptosporidium</i> spp. in Stool Samples |
| title_short | Comparison of Three Real-Time PCR Assays Targeting the SSU rRNA Gene, the COWP Gene and the DnaJ-Like Protein Gene for the Diagnosis of <i>Cryptosporidium</i> spp. in Stool Samples |
| title_sort | comparison of three real time pcr assays targeting the ssu rrna gene the cowp gene and the dnaj like protein gene for the diagnosis of i cryptosporidium i spp in stool samples |
| topic | <i>Cryptosporidium</i> spp. real-time PCR test comparison latent class analysis sensitivity specificity |
| url | https://www.mdpi.com/2076-0817/10/9/1131 |
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