Validation of Multiplex PCR and Serology Detecting IHelicobacter/I Species in Mice
High-throughput multiplexed assays are needed to simplify detection of Helicobacter species in experimental infection and routine health monitoring of laboratory mice. Therefore, fluorescent bead-based hybridization assays for Helicobacter sp. DNA and serology were developed. Multiplex PCR amplicons...
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| Veröffentlicht in: | Microorganisms (Basel) 2023-01, Vol.11 (2) |
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| creator | Butt, Julia Schmitz, Mareike Berkus, Bernhard Schmidt, Katja Höfler, Daniela |
| description | High-throughput multiplexed assays are needed to simplify detection of Helicobacter species in experimental infection and routine health monitoring of laboratory mice. Therefore, fluorescent bead-based hybridization assays for Helicobacter sp. DNA and serology were developed. Multiplex PCR amplicons (H. hepaticus, H. bilis, H. typhlonius, H. pylori, H. muridarum, H. pullorum, H. cinaedi, H. heilmanii, C. jejuni) and antibodies against H. pylori, H. hepaticus, H. bilis were assessed in naturally and experimentally infected mice, and results compared to conventional PCR. Species-specific and sensitive detection of seven Helicobacter spp. |
| doi_str_mv | 10.3390/microorganisms11020249 |
| format | Article |
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Therefore, fluorescent bead-based hybridization assays for Helicobacter sp. DNA and serology were developed. Multiplex PCR amplicons (H. hepaticus, H. bilis, H. typhlonius, H. pylori, H. muridarum, H. pullorum, H. cinaedi, H. heilmanii, C. jejuni) and antibodies against H. pylori, H. hepaticus, H. bilis were assessed in naturally and experimentally infected mice, and results compared to conventional PCR. Species-specific and sensitive detection of seven Helicobacter spp. <100 copies/PCR, and of two species <1000 copies/PCR was successfully established in the Helicobacter multiplex DNA finder. The novel assay was highly comparable with conventional PCR (kappa = 0.98, 95%CI: 0.94–1.00). Antibody detection of H. hepaticus and H. bilis showed low sensitivity (71% and 62%, respectively) and cross-reactivity in H. typhlonius-infected mice. Infection experiments showed that antibodies develop earliest two weeks after DNA detection in feces. In conclusion, detection of Helicobacter antibodies showed low sensitivity depending on the timing relative to infection. However, Helicobacter multiplex DNA finder is a sensitive and specific high-throughput assay applicable in routine health monitoring for laboratory animals.</description><identifier>ISSN: 2076-2607</identifier><identifier>EISSN: 2076-2607</identifier><identifier>DOI: 10.3390/microorganisms11020249</identifier><language>eng</language><publisher>MDPI AG</publisher><subject>Antibodies ; Antigen-antibody reactions ; Causes of ; Diseases and pests ; Genetic aspects ; Gram-negative bacteria ; Helicobacter infections ; Mice ; Microbiological research ; Risk factors ; Viral antibodies</subject><ispartof>Microorganisms (Basel), 2023-01, Vol.11 (2)</ispartof><rights>COPYRIGHT 2023 MDPI AG</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780,860,27903,27904</link.rule.ids></links><search><creatorcontrib>Butt, Julia</creatorcontrib><creatorcontrib>Schmitz, Mareike</creatorcontrib><creatorcontrib>Berkus, Bernhard</creatorcontrib><creatorcontrib>Schmidt, Katja</creatorcontrib><creatorcontrib>Höfler, Daniela</creatorcontrib><title>Validation of Multiplex PCR and Serology Detecting IHelicobacter/I Species in Mice</title><title>Microorganisms (Basel)</title><description>High-throughput multiplexed assays are needed to simplify detection of Helicobacter species in experimental infection and routine health monitoring of laboratory mice. Therefore, fluorescent bead-based hybridization assays for Helicobacter sp. DNA and serology were developed. Multiplex PCR amplicons (H. hepaticus, H. bilis, H. typhlonius, H. pylori, H. muridarum, H. pullorum, H. cinaedi, H. heilmanii, C. jejuni) and antibodies against H. pylori, H. hepaticus, H. bilis were assessed in naturally and experimentally infected mice, and results compared to conventional PCR. Species-specific and sensitive detection of seven Helicobacter spp. <100 copies/PCR, and of two species <1000 copies/PCR was successfully established in the Helicobacter multiplex DNA finder. The novel assay was highly comparable with conventional PCR (kappa = 0.98, 95%CI: 0.94–1.00). Antibody detection of H. hepaticus and H. bilis showed low sensitivity (71% and 62%, respectively) and cross-reactivity in H. typhlonius-infected mice. Infection experiments showed that antibodies develop earliest two weeks after DNA detection in feces. In conclusion, detection of Helicobacter antibodies showed low sensitivity depending on the timing relative to infection. However, Helicobacter multiplex DNA finder is a sensitive and specific high-throughput assay applicable in routine health monitoring for laboratory animals.</description><subject>Antibodies</subject><subject>Antigen-antibody reactions</subject><subject>Causes of</subject><subject>Diseases and pests</subject><subject>Genetic aspects</subject><subject>Gram-negative bacteria</subject><subject>Helicobacter infections</subject><subject>Mice</subject><subject>Microbiological research</subject><subject>Risk factors</subject><subject>Viral antibodies</subject><issn>2076-2607</issn><issn>2076-2607</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2023</creationdate><recordtype>article</recordtype><sourceid/><recordid>eNptkEtLAzEUhYMoWGr_ggRcT5t3JstSHy20KG1xWzJ5DJFMUiYj6L93QBddeO_iHA7fPYsLwD1Gc0oVWnTB9Dn3rU6hdAVjRBBh6gpMCJKiIgLJ6wt_C2alfKBxFKY1xxOwf9cxWD2EnGD2cPcZh3CO7gu-rfZQJwsPrs8xt9_w0Q3ODCG1cLN2MZjcaDO4frGBh7MzwRUYEtwF4-7AjdexuNmfTsHx-em4Wlfb15fNarmtWiFZJT11HCHSMFszrLgQDfbcW9sYzpGoPda1YsrLWkgimBo5bhBlVhGutBB0Ch5-a1sd3Skkn4demy4Uc1pKRmpFpWIjNf-HGte68XU5OR_G_OLgBw8oY-4</recordid><startdate>20230101</startdate><enddate>20230101</enddate><creator>Butt, Julia</creator><creator>Schmitz, Mareike</creator><creator>Berkus, Bernhard</creator><creator>Schmidt, Katja</creator><creator>Höfler, Daniela</creator><general>MDPI AG</general><scope/></search><sort><creationdate>20230101</creationdate><title>Validation of Multiplex PCR and Serology Detecting IHelicobacter/I Species in Mice</title><author>Butt, Julia ; Schmitz, Mareike ; Berkus, Bernhard ; Schmidt, Katja ; Höfler, Daniela</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-g674-7f3e5002b4d8419566b1f5fddbc55068f1a8949f78672649b4d5c034d9259a663</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2023</creationdate><topic>Antibodies</topic><topic>Antigen-antibody reactions</topic><topic>Causes of</topic><topic>Diseases and pests</topic><topic>Genetic aspects</topic><topic>Gram-negative bacteria</topic><topic>Helicobacter infections</topic><topic>Mice</topic><topic>Microbiological research</topic><topic>Risk factors</topic><topic>Viral antibodies</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Butt, Julia</creatorcontrib><creatorcontrib>Schmitz, Mareike</creatorcontrib><creatorcontrib>Berkus, Bernhard</creatorcontrib><creatorcontrib>Schmidt, Katja</creatorcontrib><creatorcontrib>Höfler, Daniela</creatorcontrib><jtitle>Microorganisms (Basel)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Butt, Julia</au><au>Schmitz, Mareike</au><au>Berkus, Bernhard</au><au>Schmidt, Katja</au><au>Höfler, Daniela</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Validation of Multiplex PCR and Serology Detecting IHelicobacter/I Species in Mice</atitle><jtitle>Microorganisms (Basel)</jtitle><date>2023-01-01</date><risdate>2023</risdate><volume>11</volume><issue>2</issue><issn>2076-2607</issn><eissn>2076-2607</eissn><abstract>High-throughput multiplexed assays are needed to simplify detection of Helicobacter species in experimental infection and routine health monitoring of laboratory mice. Therefore, fluorescent bead-based hybridization assays for Helicobacter sp. DNA and serology were developed. Multiplex PCR amplicons (H. hepaticus, H. bilis, H. typhlonius, H. pylori, H. muridarum, H. pullorum, H. cinaedi, H. heilmanii, C. jejuni) and antibodies against H. pylori, H. hepaticus, H. bilis were assessed in naturally and experimentally infected mice, and results compared to conventional PCR. Species-specific and sensitive detection of seven Helicobacter spp. <100 copies/PCR, and of two species <1000 copies/PCR was successfully established in the Helicobacter multiplex DNA finder. The novel assay was highly comparable with conventional PCR (kappa = 0.98, 95%CI: 0.94–1.00). Antibody detection of H. hepaticus and H. bilis showed low sensitivity (71% and 62%, respectively) and cross-reactivity in H. typhlonius-infected mice. Infection experiments showed that antibodies develop earliest two weeks after DNA detection in feces. In conclusion, detection of Helicobacter antibodies showed low sensitivity depending on the timing relative to infection. However, Helicobacter multiplex DNA finder is a sensitive and specific high-throughput assay applicable in routine health monitoring for laboratory animals.</abstract><pub>MDPI AG</pub><doi>10.3390/microorganisms11020249</doi></addata></record> |
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| source | DOAJ Directory of Open Access Journals; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; PubMed Central Open Access; MDPI - Multidisciplinary Digital Publishing Institute; PubMed Central |
| subjects | Antibodies Antigen-antibody reactions Causes of Diseases and pests Genetic aspects Gram-negative bacteria Helicobacter infections Mice Microbiological research Risk factors Viral antibodies |
| title | Validation of Multiplex PCR and Serology Detecting IHelicobacter/I Species in Mice |
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