Molecular detection and speciation of pathogenic <it>Leptospira </it>spp. in blood from patients with culture-negative leptospirosis
<p>Abstract</p> <p>Background</p> <p>Pathogenic <it>Leptospira </it>spp. present in the blood of patients with leptospirosis during the first week of symptoms can be detected using culture or PCR. A proportion of patients who are positive by PCR are negative...
| Main Authors: | , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2011-12-01
|
| Series: | BMC Infectious Diseases |
| Online Access: | http://www.biomedcentral.com/1471-2334/11/338 |
| _version_ | 1811280814267170816 |
|---|---|
| author | Boonsilp Siriphan Thaipadungpanit Janjira Amornchai Premjit Wuthiekanun Vanaporn Chierakul Wirongrong Limmathurotsakul Direk Day Nicholas P Peacock Sharon J |
| author_facet | Boonsilp Siriphan Thaipadungpanit Janjira Amornchai Premjit Wuthiekanun Vanaporn Chierakul Wirongrong Limmathurotsakul Direk Day Nicholas P Peacock Sharon J |
| author_sort | Boonsilp Siriphan |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p>Pathogenic <it>Leptospira </it>spp. present in the blood of patients with leptospirosis during the first week of symptoms can be detected using culture or PCR. A proportion of patients who are positive by PCR are negative by culture. <it>Leptospira </it>spp. are fastidious bacteria, and we hypothesized that a false-negative culture result may represent infection with a distinct bacterial subset that fail to grow in standard culture medium.</p> <p>Methods</p> <p>We evaluated our hypothesis during a prospective study of 418 consecutive patients presenting to a hospital in northeast Thailand with an acute febrile illness. Admission blood samples were taken for <it>Leptospira </it>culture and PCR. A single tube nested PCR that amplified a region of the <it>rrs </it>gene was developed and applied, amplicons sequenced and a phylogenetic tree reconstructed.</p> <p>Results</p> <p>39/418 (9%) patients were culture-positive for <it>Leptospira </it>spp., and 81/418 (19%) patients were culture-negative but <it>rrs </it>PCR-positive. The species associated with culture-positive leptospirosis (37 <it>L. interrogans </it>and 2 <it>L. borgpetersenii</it>) were comparable to those associated with culture-negative, PCR-positive leptospirosis (76 <it>L. interrogans</it>, 4 <it>L. borgpetersenii</it>, 1 unidentified, possibly new species).</p> <p>Conclusion</p> <p>Molecular speciation failed to identify a unique bacterial subset in patients with culture-negative, PCR-positive leptospirosis. The rate of false-negative culture was high, and we speculate that antibiotic pre-treatment is the most likely explanation for this.</p> |
| first_indexed | 2024-04-13T01:21:34Z |
| format | Article |
| id | doaj.art-ee9fa431caac4fd595a37e13e2368b3f |
| institution | Directory Open Access Journal |
| issn | 1471-2334 |
| language | English |
| last_indexed | 2024-04-13T01:21:34Z |
| publishDate | 2011-12-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Infectious Diseases |
| spelling | doaj.art-ee9fa431caac4fd595a37e13e2368b3f2022-12-22T03:08:45ZengBMCBMC Infectious Diseases1471-23342011-12-0111133810.1186/1471-2334-11-338Molecular detection and speciation of pathogenic <it>Leptospira </it>spp. in blood from patients with culture-negative leptospirosisBoonsilp SiriphanThaipadungpanit JanjiraAmornchai PremjitWuthiekanun VanapornChierakul WirongrongLimmathurotsakul DirekDay Nicholas PPeacock Sharon J<p>Abstract</p> <p>Background</p> <p>Pathogenic <it>Leptospira </it>spp. present in the blood of patients with leptospirosis during the first week of symptoms can be detected using culture or PCR. A proportion of patients who are positive by PCR are negative by culture. <it>Leptospira </it>spp. are fastidious bacteria, and we hypothesized that a false-negative culture result may represent infection with a distinct bacterial subset that fail to grow in standard culture medium.</p> <p>Methods</p> <p>We evaluated our hypothesis during a prospective study of 418 consecutive patients presenting to a hospital in northeast Thailand with an acute febrile illness. Admission blood samples were taken for <it>Leptospira </it>culture and PCR. A single tube nested PCR that amplified a region of the <it>rrs </it>gene was developed and applied, amplicons sequenced and a phylogenetic tree reconstructed.</p> <p>Results</p> <p>39/418 (9%) patients were culture-positive for <it>Leptospira </it>spp., and 81/418 (19%) patients were culture-negative but <it>rrs </it>PCR-positive. The species associated with culture-positive leptospirosis (37 <it>L. interrogans </it>and 2 <it>L. borgpetersenii</it>) were comparable to those associated with culture-negative, PCR-positive leptospirosis (76 <it>L. interrogans</it>, 4 <it>L. borgpetersenii</it>, 1 unidentified, possibly new species).</p> <p>Conclusion</p> <p>Molecular speciation failed to identify a unique bacterial subset in patients with culture-negative, PCR-positive leptospirosis. The rate of false-negative culture was high, and we speculate that antibiotic pre-treatment is the most likely explanation for this.</p>http://www.biomedcentral.com/1471-2334/11/338 |
| spellingShingle | Boonsilp Siriphan Thaipadungpanit Janjira Amornchai Premjit Wuthiekanun Vanaporn Chierakul Wirongrong Limmathurotsakul Direk Day Nicholas P Peacock Sharon J Molecular detection and speciation of pathogenic <it>Leptospira </it>spp. in blood from patients with culture-negative leptospirosis BMC Infectious Diseases |
| title | Molecular detection and speciation of pathogenic <it>Leptospira </it>spp. in blood from patients with culture-negative leptospirosis |
| title_full | Molecular detection and speciation of pathogenic <it>Leptospira </it>spp. in blood from patients with culture-negative leptospirosis |
| title_fullStr | Molecular detection and speciation of pathogenic <it>Leptospira </it>spp. in blood from patients with culture-negative leptospirosis |
| title_full_unstemmed | Molecular detection and speciation of pathogenic <it>Leptospira </it>spp. in blood from patients with culture-negative leptospirosis |
| title_short | Molecular detection and speciation of pathogenic <it>Leptospira </it>spp. in blood from patients with culture-negative leptospirosis |
| title_sort | molecular detection and speciation of pathogenic it leptospira it spp in blood from patients with culture negative leptospirosis |
| url | http://www.biomedcentral.com/1471-2334/11/338 |
| work_keys_str_mv | AT boonsilpsiriphan moleculardetectionandspeciationofpathogenicitleptospiraitsppinbloodfrompatientswithculturenegativeleptospirosis AT thaipadungpanitjanjira moleculardetectionandspeciationofpathogenicitleptospiraitsppinbloodfrompatientswithculturenegativeleptospirosis AT amornchaipremjit moleculardetectionandspeciationofpathogenicitleptospiraitsppinbloodfrompatientswithculturenegativeleptospirosis AT wuthiekanunvanaporn moleculardetectionandspeciationofpathogenicitleptospiraitsppinbloodfrompatientswithculturenegativeleptospirosis AT chierakulwirongrong moleculardetectionandspeciationofpathogenicitleptospiraitsppinbloodfrompatientswithculturenegativeleptospirosis AT limmathurotsakuldirek moleculardetectionandspeciationofpathogenicitleptospiraitsppinbloodfrompatientswithculturenegativeleptospirosis AT daynicholasp moleculardetectionandspeciationofpathogenicitleptospiraitsppinbloodfrompatientswithculturenegativeleptospirosis AT peacocksharonj moleculardetectionandspeciationofpathogenicitleptospiraitsppinbloodfrompatientswithculturenegativeleptospirosis |