ES2887019T3 - Procedimiento para secretar extracelularmente una proteína específica del complejo de mycobacterium tuberculosis - Google Patents
Procedimiento para secretar extracelularmente una proteína específica del complejo de mycobacterium tuberculosis Download PDFInfo
- Publication number
- ES2887019T3 ES2887019T3 ES19150248T ES19150248T ES2887019T3 ES 2887019 T3 ES2887019 T3 ES 2887019T3 ES 19150248 T ES19150248 T ES 19150248T ES 19150248 T ES19150248 T ES 19150248T ES 2887019 T3 ES2887019 T3 ES 2887019T3
- Authority
- ES
- Spain
- Prior art keywords
- mycobacterium tuberculosis
- tuberculosis complex
- biological sample
- extracellularly
- sample
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 241001302239 Mycobacterium tuberculosis complex Species 0.000 title claims abstract description 69
- 238000000034 method Methods 0.000 title claims abstract description 38
- 102000004169 proteins and genes Human genes 0.000 title claims abstract description 26
- 108090000623 proteins and genes Proteins 0.000 title claims abstract description 26
- 230000003248 secreting effect Effects 0.000 title claims abstract description 12
- 239000012472 biological sample Substances 0.000 claims abstract description 51
- 238000010438 heat treatment Methods 0.000 claims abstract description 51
- 101100291912 Mycobacterium bovis (strain ATCC BAA-935 / AF2122/97) mpb64 gene Proteins 0.000 claims description 38
- 239000000523 sample Substances 0.000 claims description 34
- 230000028327 secretion Effects 0.000 claims description 15
- 102000040739 Secretory proteins Human genes 0.000 claims description 13
- 108091058545 Secretory proteins Proteins 0.000 claims description 13
- 101150014428 mpt64 gene Proteins 0.000 claims description 12
- 239000002904 solvent Substances 0.000 claims description 12
- 238000003556 assay Methods 0.000 claims description 10
- 238000010324 immunological assay Methods 0.000 claims description 10
- 239000003795 chemical substances by application Substances 0.000 claims description 4
- 238000003018 immunoassay Methods 0.000 claims description 4
- 210000002700 urine Anatomy 0.000 claims description 4
- 238000002965 ELISA Methods 0.000 claims description 3
- 208000002151 Pleural effusion Diseases 0.000 claims description 2
- 210000004369 blood Anatomy 0.000 claims description 2
- 239000008280 blood Substances 0.000 claims description 2
- 210000000621 bronchi Anatomy 0.000 claims description 2
- 210000001175 cerebrospinal fluid Anatomy 0.000 claims description 2
- 210000003608 fece Anatomy 0.000 claims description 2
- 239000012530 fluid Substances 0.000 claims description 2
- 210000004051 gastric juice Anatomy 0.000 claims description 2
- 210000004072 lung Anatomy 0.000 claims description 2
- 210000001519 tissue Anatomy 0.000 claims description 2
- 230000001580 bacterial effect Effects 0.000 description 50
- 239000000243 solution Substances 0.000 description 24
- 238000001514 detection method Methods 0.000 description 19
- 238000002835 absorbance Methods 0.000 description 16
- 208000015181 infectious disease Diseases 0.000 description 16
- 206010036790 Productive cough Diseases 0.000 description 13
- 210000004027 cell Anatomy 0.000 description 13
- 210000003802 sputum Anatomy 0.000 description 13
- 208000024794 sputum Diseases 0.000 description 13
- 239000007788 liquid Substances 0.000 description 12
- 201000008827 tuberculosis Diseases 0.000 description 12
- 238000011156 evaluation Methods 0.000 description 10
- 238000012360 testing method Methods 0.000 description 8
- 239000000126 substance Substances 0.000 description 7
- 241000894006 Bacteria Species 0.000 description 6
- 241000187479 Mycobacterium tuberculosis Species 0.000 description 6
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 6
- 238000012258 culturing Methods 0.000 description 6
- 208000003322 Coinfection Diseases 0.000 description 5
- 230000003321 amplification Effects 0.000 description 5
- 238000003199 nucleic acid amplification method Methods 0.000 description 5
- 102000039446 nucleic acids Human genes 0.000 description 5
- 108020004707 nucleic acids Proteins 0.000 description 5
- 150000007523 nucleic acids Chemical class 0.000 description 5
- 241001467552 Mycobacterium bovis BCG Species 0.000 description 4
- 230000001900 immune effect Effects 0.000 description 4
- 230000003287 optical effect Effects 0.000 description 4
- 230000001737 promoting effect Effects 0.000 description 4
- 239000007787 solid Substances 0.000 description 4
- 239000000427 antigen Substances 0.000 description 3
- 102000036639 antigens Human genes 0.000 description 3
- 108091007433 antigens Proteins 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 230000002779 inactivation Effects 0.000 description 3
- 239000003550 marker Substances 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 description 3
- 229920000053 polysorbate 80 Polymers 0.000 description 3
- 230000000007 visual effect Effects 0.000 description 3
- 208000035473 Communicable disease Diseases 0.000 description 2
- PWKSKIMOESPYIA-BYPYZUCNSA-N L-N-acetyl-Cysteine Chemical compound CC(=O)N[C@@H](CS)C(O)=O PWKSKIMOESPYIA-BYPYZUCNSA-N 0.000 description 2
- 241000186367 Mycobacterium avium Species 0.000 description 2
- 241000186366 Mycobacterium bovis Species 0.000 description 2
- 239000003638 chemical reducing agent Substances 0.000 description 2
- 238000007796 conventional method Methods 0.000 description 2
- 238000003745 diagnosis Methods 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 208000027531 mycobacterial infectious disease Diseases 0.000 description 2
- 239000008055 phosphate buffer solution Substances 0.000 description 2
- 230000005180 public health Effects 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 101710166488 6 kDa early secretory antigenic target Proteins 0.000 description 1
- 102000009027 Albumins Human genes 0.000 description 1
- 108010088751 Albumins Proteins 0.000 description 1
- 241000283690 Bos taurus Species 0.000 description 1
- 102000016938 Catalase Human genes 0.000 description 1
- 108010053835 Catalase Proteins 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- 241001529936 Murinae Species 0.000 description 1
- 206010062207 Mycobacterial infection Diseases 0.000 description 1
- 241001467553 Mycobacterium africanum Species 0.000 description 1
- 101100131080 Mycobacterium bovis (strain ATCC BAA-935 / AF2122/97) mpb70 gene Proteins 0.000 description 1
- 241000186363 Mycobacterium kansasii Species 0.000 description 1
- 241000187492 Mycobacterium marinum Species 0.000 description 1
- 241000187919 Mycobacterium microti Species 0.000 description 1
- 101001057048 Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv) ESAT-6-like protein EsxB Proteins 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 1
- 206010044756 Tuberculous infections Diseases 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 239000013068 control sample Substances 0.000 description 1
- 230000009089 cytolysis Effects 0.000 description 1
- 239000003398 denaturant Substances 0.000 description 1
- 239000008121 dextrose Substances 0.000 description 1
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 230000002458 infectious effect Effects 0.000 description 1
- 230000000977 initiatory effect Effects 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 239000002953 phosphate buffered saline Substances 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000000241 respiratory effect Effects 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 239000004094 surface-active agent Substances 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 238000007669 thermal treatment Methods 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
- G01N33/56911—Bacteria
- G01N33/5695—Mycobacteria
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
- G01N33/56911—Bacteria
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Immunology (AREA)
- Engineering & Computer Science (AREA)
- Urology & Nephrology (AREA)
- Hematology (AREA)
- Biomedical Technology (AREA)
- Chemical & Material Sciences (AREA)
- Molecular Biology (AREA)
- Medicinal Chemistry (AREA)
- Biochemistry (AREA)
- Cell Biology (AREA)
- Pathology (AREA)
- Biotechnology (AREA)
- Food Science & Technology (AREA)
- General Physics & Mathematics (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Microbiology (AREA)
- General Health & Medical Sciences (AREA)
- Virology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Sampling And Sample Adjustment (AREA)
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2012086566 | 2012-04-05 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| ES2887019T3 true ES2887019T3 (es) | 2021-12-21 |
Family
ID=49300600
Family Applications (3)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| ES19150248T Active ES2887019T3 (es) | 2012-04-05 | 2013-04-04 | Procedimiento para secretar extracelularmente una proteína específica del complejo de mycobacterium tuberculosis |
| ES13772467T Active ES2751357T3 (es) | 2012-04-05 | 2013-04-04 | Procedimiento de detección inmunológica del complejo Mycobacterium tuberculosis |
| ES16166865T Active ES2746565T3 (es) | 2012-04-05 | 2013-04-04 | Procedimiento y kit para la detección inmunológica de complejo de Mycobacterium tuberculosis |
Family Applications After (2)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| ES13772467T Active ES2751357T3 (es) | 2012-04-05 | 2013-04-04 | Procedimiento de detección inmunológica del complejo Mycobacterium tuberculosis |
| ES16166865T Active ES2746565T3 (es) | 2012-04-05 | 2013-04-04 | Procedimiento y kit para la detección inmunológica de complejo de Mycobacterium tuberculosis |
Country Status (9)
| Country | Link |
|---|---|
| US (3) | US10823730B2 (https=) |
| EP (3) | EP2835646B1 (https=) |
| JP (2) | JP5612778B2 (https=) |
| CN (3) | CN104246505B (https=) |
| BR (1) | BR112014024574B1 (https=) |
| ES (3) | ES2887019T3 (https=) |
| IN (1) | IN2014DN08246A (https=) |
| RU (1) | RU2657581C2 (https=) |
| WO (1) | WO2013151122A1 (https=) |
Families Citing this family (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP2835646B1 (en) | 2012-04-05 | 2019-07-17 | Tauns Co., Ltd. | Method for immunological detection of mycobacterium tuberculosis complex |
| WO2015164617A1 (en) * | 2014-04-24 | 2015-10-29 | Somalogic, Inc. | Tuberculosis biomarkers in urine and uses thereof |
| RU2594063C1 (ru) * | 2015-04-27 | 2016-08-10 | Федеральное государственное бюджетное научное учреждение "Центральный научно-исследовательский институт туберкулёза" | Способ диагностики микобактерий туберкулёза |
| CN115181781A (zh) | 2015-11-02 | 2022-10-14 | 拜奥法尔诊断有限责任公司 | 困难样品类型的样品制备 |
| PL233267B1 (pl) * | 2017-10-16 | 2019-09-30 | Politechnika Warszawska | Sposób detekcji bakterii gruźlicy |
| JP6989428B2 (ja) * | 2018-03-23 | 2022-01-05 | アークレイ株式会社 | 前処理剤、前処理キット、抗原検査キット及び抗原検査方法 |
| US20230193346A1 (en) * | 2020-05-06 | 2023-06-22 | Korea Atomic Energy Research Institute | Method for detecting mycobacterium tuberculosis |
Family Cites Families (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE3700049A1 (de) * | 1987-01-02 | 1988-07-14 | Biotest Serum Institut Gmbh | Verfahren zum immunologischen nachweis von mykobakterien im sputum sowie monoklonale antikoerper zur durchfuehrung des verfahrens |
| JPH07110332A (ja) * | 1993-10-12 | 1995-04-25 | Tetsuo Tomiyama | 結核菌の検査試薬及び検出方法 |
| SE9600949D0 (sv) | 1996-03-12 | 1996-03-12 | Stefan Svenson | Method of diagnosing a mycobacterial disease and immunoassay kit |
| JP4269006B2 (ja) | 1997-09-30 | 2009-05-27 | 株式会社ビーエル | 結核菌検出法 |
| JP2002062299A (ja) * | 2000-08-24 | 2002-02-28 | Kyowa Medex Co Ltd | 結核菌の検出方法及び試薬 |
| JP2004166564A (ja) * | 2002-11-19 | 2004-06-17 | Bl:Kk | モノクローナル抗体および結核症診断法 |
| JP4419020B2 (ja) * | 2005-02-25 | 2010-02-24 | 長庚大學 | 体液中の結核菌抗原を検出する方法 |
| CN101336375A (zh) * | 2005-12-26 | 2008-12-31 | 株式会社比尔生命 | 用于检测流感a型病毒的强毒株的方法 |
| JP4478658B2 (ja) * | 2006-03-31 | 2010-06-09 | 株式会社ビーエル | 結核菌検出用キット |
| US8541179B2 (en) * | 2007-12-28 | 2013-09-24 | Bl Co., Ltd. | Immunodetection assay for Mycobacterium tuberculosis complex |
| EP2835646B1 (en) * | 2012-04-05 | 2019-07-17 | Tauns Co., Ltd. | Method for immunological detection of mycobacterium tuberculosis complex |
-
2013
- 2013-04-04 EP EP13772467.0A patent/EP2835646B1/en active Active
- 2013-04-04 EP EP19150248.3A patent/EP3499237B1/en active Active
- 2013-04-04 CN CN201380018968.0A patent/CN104246505B/zh active Active
- 2013-04-04 CN CN201610009079.3A patent/CN105403700B/zh active Active
- 2013-04-04 IN IN8246DEN2014 patent/IN2014DN08246A/en unknown
- 2013-04-04 RU RU2014144283A patent/RU2657581C2/ru active
- 2013-04-04 CN CN201610245191.7A patent/CN105784999B/zh active Active
- 2013-04-04 EP EP16166865.2A patent/EP3093667B1/en active Active
- 2013-04-04 ES ES19150248T patent/ES2887019T3/es active Active
- 2013-04-04 ES ES13772467T patent/ES2751357T3/es active Active
- 2013-04-04 ES ES16166865T patent/ES2746565T3/es active Active
- 2013-04-04 BR BR112014024574-6A patent/BR112014024574B1/pt active IP Right Grant
- 2013-04-04 WO PCT/JP2013/060301 patent/WO2013151122A1/ja not_active Ceased
- 2013-04-04 US US14/390,706 patent/US10823730B2/en active Active
- 2013-04-04 JP JP2013539828A patent/JP5612778B2/ja active Active
-
2014
- 2014-09-04 JP JP2014179856A patent/JP6216299B2/ja active Active
-
2016
- 2016-04-28 US US15/141,384 patent/US10883988B2/en active Active
-
2018
- 2018-05-16 US US15/980,920 patent/US10830769B2/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| EP2835646A4 (en) | 2015-12-02 |
| CN105403700A (zh) | 2016-03-16 |
| JP6216299B2 (ja) | 2017-10-18 |
| IN2014DN08246A (https=) | 2015-07-10 |
| EP2835646B1 (en) | 2019-07-17 |
| EP3499237B1 (en) | 2021-06-09 |
| US10883988B2 (en) | 2021-01-05 |
| EP2835646A1 (en) | 2015-02-11 |
| CN104246505B (zh) | 2016-08-24 |
| US20180259519A1 (en) | 2018-09-13 |
| RU2014144283A (ru) | 2016-05-27 |
| CN105784999A (zh) | 2016-07-20 |
| US10823730B2 (en) | 2020-11-03 |
| ES2751357T3 (es) | 2020-03-31 |
| JPWO2013151122A1 (ja) | 2015-12-17 |
| EP3499237A1 (en) | 2019-06-19 |
| EP3093667B1 (en) | 2019-07-10 |
| JP5612778B2 (ja) | 2014-10-22 |
| EP3093667A1 (en) | 2016-11-16 |
| CN105403700B (zh) | 2018-03-27 |
| US10830769B2 (en) | 2020-11-10 |
| ES2746565T3 (es) | 2020-03-06 |
| WO2013151122A1 (ja) | 2013-10-10 |
| JP2014232117A (ja) | 2014-12-11 |
| CN104246505A (zh) | 2014-12-24 |
| US20150064729A1 (en) | 2015-03-05 |
| US20160238599A1 (en) | 2016-08-18 |
| CN105784999B (zh) | 2018-05-22 |
| RU2657581C2 (ru) | 2018-06-14 |
| BR112014024574B1 (pt) | 2020-10-27 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| ES2887019T3 (es) | Procedimiento para secretar extracelularmente una proteína específica del complejo de mycobacterium tuberculosis | |
| Lilenbaum et al. | Detection of Leptospira spp. in semen and vaginal fluids of goats and sheep by polymerase chain reaction | |
| Yaakob et al. | Leptospirosis: recent incidents and available diagnostics–a review | |
| Pfyffer et al. | Mycobacterium: general characteristics, laboratory detection, and staining procedures | |
| Al-Orry et al. | A review of laboratory diagnosis and treatment of leptospirosis | |
| Stich et al. | Evaluation of an automated system for non-radiometric detection of Mycobacterium avium paratuberculosis in bovine feces | |
| ES2280630T3 (es) | Dispositivos para recoger y preparar especimenes para la deteccion de microbacterias y sus antigenos. | |
| Safiulah et al. | Laboratory methods for diagnosing leptospirosis: a review | |
| Hatem et al. | Surveillance of bovine leptospirosis: isolation and serodiagnosis | |
| Roseliza et al. | Diagnosis of paratuberculosis by microbiological culture in Veterinary Research Institute from 2001 to 2018. | |
| HK1224737B (en) | Method and kit for immunological detection of mycobacterium tuberculosis complex | |
| HK1224737A1 (en) | Method and kit for immunological detection of mycobacterium tuberculosis complex | |
| HK1224737A (en) | Method and kit for immunological detection of mycobacterium tuberculosis complex | |
| Usharani et al. | Molecular epidemiology of female genital tuberculosis leading to infertility | |
| Ali et al. | Identification of Bovine and human tuberculosis in AFB positive and negative patients | |
| Verma | NONTUBERCULOUS MYCOBACTERIA (NTM) | |
| Russo et al. | Q fever | |
| Minda Asfaw Geresu et al. | A review on diagnostic methods of brucellosis. | |
| SUBI | REAL-TIME PCR BASED DETECTION OF LEPTOSPIRES IN BLOOD AND URINE OF DOGS | |
| Subramaniyan et al. | Seroepidemiological review of leptospirosis and its co-infection between 2007 to 2009 in Chennai, Tamil Nadu--a doctoral thesis report | |
| Balakrishnan et al. | Issues in the diagnosis of leptospirosis. | |
| Denton | Digital Commons@ University o f South Florid |