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Article ; Online: Stepwise Protocol for Cytospin-enhanced Smearing for Scraped Corneal Epithelial Cells.

Jeyalatha, Mani V / Malathi, Jambulingam / Madhavan, Hajib N

Applied immunohistochemistry & molecular morphology : AIMM

2016 Volume 24, Issue 1, Page(s) 71–73

Abstract: Proteins and antigens present on the cell surface are usually determined by immunofluorescence staining. Uniform distribution of cells is required to appreciate the presence of surface proteins. Improper smearing or crushing of the corneal epithelial ... ...

Abstract	Proteins and antigens present on the cell surface are usually determined by immunofluorescence staining. Uniform distribution of cells is required to appreciate the presence of surface proteins. Improper smearing or crushing of the corneal epithelial cells can potentially destroy the cellular integrity. Thus a simplified, systemic method was designed to smear the cells scraped from the cornea. The procedure includes trypsinisation for dissociation of corneal epithelial cells and cytospinning for concentrating the cells in a smear. The standardized protocol was found to be efficient in maintaining the integrity of the corneal epithelial cells and also the distribution of the cells in the smear.
MeSH term(s)	Biomarkers/metabolism ; Cell Adhesion ; Cell Separation/methods ; Centrifugation ; Cornea/cytology ; Cornea/metabolism ; Epithelial Cells/cytology ; Epithelial Cells/metabolism ; Fluorescent Antibody Technique ; Gene Expression ; Humans ; Microscopy, Fluorescence ; Receptors, TNF-Related Apoptosis-Inducing Ligand/genetics ; Trypsin/chemistry
Chemical Substances	Biomarkers ; Receptors, TNF-Related Apoptosis-Inducing Ligand ; TNFRSF10A protein, human ; Trypsin (EC 3.4.21.4)
Language	English
Publishing date	2016-01
Publishing country	United States
Document type	Journal Article
ZDB-ID	1473273-7
ISSN	1533-4058 ; 1062-3345 ; 1541-2016
ISSN (online)	1533-4058
ISSN	1062-3345 ; 1541-2016
DOI	10.1097/PAI.0000000000000168
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Zs.A 3783: Show issues

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Article ; Online: Unveiling the ocular battlefield: Insights into Pseudomonas aeruginosa virulence factors and their implications for multidrug resistance.

Murugan, Nandagopal / Krishnamoorthy, Rajapandiyan / Khan, Javed Masood / Gatasheh, Mansour K / Malathi, Jambulingam / Madhavan, Hajib Narahari Rao / Ramalingam, Gopinath / Jayaramana, Selvaraj

International journal of biological macromolecules

2024 Volume 267, Issue Pt 2, Page(s) 131677

Abstract: The research investigates the virulence factors of Pseudomonas aeruginosa (P. aeruginosa), a pathogen known for its ability to cause human infections by releasing various exoenzymes and virulence factors. Particularly relevant in ocular infections, where ...

Abstract	The research investigates the virulence factors of Pseudomonas aeruginosa (P. aeruginosa), a pathogen known for its ability to cause human infections by releasing various exoenzymes and virulence factors. Particularly relevant in ocular infections, where tissue degeneration can occur, even after bacterial growth has ceased due to the potential role of secreted proteins/enzymes. Clinical isolates of P. aeruginosa, both ocular (146) and non-ocular (54), were examined to determine the frequency and mechanism of virulence factors. Phenotypic characterization revealed the production of alginate, biofilm, phospholipase C, and alkaline protease, while genotypic testing using internal uniplex PCR identified the presence of Exo U, S, T, Y, and LasB genes. Results showed a significant prevalence of Exo U and Y genes in ocular isolates, a finding unique to Indian studies. Additionally, the study noted that ocular isolates often contained all four secretomes, suggesting a potential link between these factors and ocular infections. These findings contribute to understanding the pathogenesis of P. aeruginosa infections, particularly in ocular contexts, and highlights the importance of comprehensive virulence factor analysis in clinical settings.
Language	English
Publishing date	2024-04-17
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	282732-3
ISSN	1879-0003 ; 0141-8130
ISSN (online)	1879-0003
ISSN	0141-8130
DOI	10.1016/j.ijbiomac.2024.131677
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Zs.B 2374: Show issues

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Article ; Online: Therapeutic Effect of Corneal Crosslinking on Fungal Keratitis: Efficacy of Corneal Collagen Crosslinking as an Adjuvant Therapy for Fungal Keratitis in a Tertiary Eye Hospital in South India.

Jeyalatha Mani, Vimalin / Parthasarathy, Durgadevi / Padmanabhan, Prema / Narayanan, Niveditha / Lakshmipathy, Meena / Pachayappan, Saravana Kumar / Jayavel, Padmapriya / Therese, Kulandhai Lily / Rao Madhavan, Hajib Narahari / Jambulingam, Malathi

Ocular immunology and inflammation

2020 Volume 29, Issue 7-8, Page(s) 1648–1655

Abstract: ... keratitis patients were recruited and randomized into two groups: group 1 (n= 11, standard antifungal ... group 2 (n=9, corneal collagen crosslinking with standard antifungal). Corneal scraping and tear samples ...

Abstract	Purpose: To evaluate the efficacy of CXL in treating fungal keratitis as an adjuvant therapy. Methods: Detailed clinical examination microbiological investigation was performed. Twenty fungal keratitis patients were recruited and randomized into two groups: group 1 (n= 11, standard antifungal), group 2 (n=9, corneal collagen crosslinking with standard antifungal). Corneal scraping and tear samples collected were subjected to real-time PCR targeting ITS, TLR analysis and cytokine analysis. Results: The mean time for complete resolution of ulcer for group 2 was significantly shorter compared to group 1 and the final mean BCVA was better for group 2. Expression of IL-1β, IL-8, IFN-γ significantly decreased immediately post CXL in group 2 patients. Significant downregulation of TLR 6, TLR-3, TLR-4 was observed 3-days post CXL compared to group 1 patients. Conclusion: Adjuvant effect of CXL was significant in treating fungal keratitis compared to standalone antifungal treatment.
MeSH term(s)	Adult ; Antifungal Agents/therapeutic use ; Collagen/metabolism ; Combined Modality Therapy ; Corneal Stroma/drug effects ; Corneal Stroma/metabolism ; Corneal Ulcer/drug therapy ; Corneal Ulcer/metabolism ; Corneal Ulcer/microbiology ; Cross-Linking Reagents/therapeutic use ; Cytokines/metabolism ; Eye Infections, Fungal/drug therapy ; Eye Infections, Fungal/metabolism ; Eye Infections, Fungal/microbiology ; Female ; Humans ; India ; Male ; Middle Aged ; Ophthalmology ; Photochemotherapy/methods ; Photosensitizing Agents/therapeutic use ; Riboflavin/therapeutic use ; Tertiary Care Centers ; Toll-Like Receptors/metabolism ; Treatment Outcome ; Ultraviolet Rays
Chemical Substances	Antifungal Agents ; Cross-Linking Reagents ; Cytokines ; Photosensitizing Agents ; Toll-Like Receptors ; Collagen (9007-34-5) ; Riboflavin (TLM2976OFR)
Language	English
Publishing date	2020-07-09
Publishing country	England
Document type	Journal Article
ZDB-ID	1193873-0
ISSN	1744-5078 ; 0927-3948
ISSN (online)	1744-5078
ISSN	0927-3948
DOI	10.1080/09273948.2020.1770296
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Zs.A 4004: Show issues

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Article: Diagnostic utility of polymerase chain reaction on intraocular specimens to establish the etiology of infectious endophthalmitis.

Sowmya, Parameswaran / Madhavan, Hajib N

European journal of ophthalmology

2009 Volume 19, Issue 5, Page(s) 812–817

Abstract: Purpose: To evaluate the utility of polymerase chain reaction (PCR) on intraocular clinical specimens (aqueous humor [AH] and vitreous fluid [VF]) as an etiologic diagnostic tool relative to microbiological culture methods in infectious endophthalmitis.! ...

Abstract	Purpose: To evaluate the utility of polymerase chain reaction (PCR) on intraocular clinical specimens (aqueous humor [AH] and vitreous fluid [VF]) as an etiologic diagnostic tool relative to microbiological culture methods in infectious endophthalmitis. Methods: Conventional bacterial and mycologic cultures and PCR for eubacterial and panfungal genomes were applied for etiologic diagnosis on pairs of AH and VF obtained from 72 patients with clinically established infectious endophthalmitis. Results: Based on cultures, an infectious etiology was established in 27 (37.5%) of 72 patients. PCR detected infectious etiology in all 72 patients. PCR increased the clinical sensitivity over culture by 62.5% (p<0.0001, McNemar test). The frequency of culture positivity, single infections, and polymicrobial infection varied significantly among the types of endophthalmitis (p<0.0001, chi-square test). PCR detected an infectious etiology in 48 patients and polymicrobial infection in 24 patients. An etiology was established by PCR on 56 (77.8%) AH and 65 (90.3%) VF of the 72 patients and this difference had no statistical significance. Conclusions: PCR on intraocular specimens as an etiologic diagnostic tool has been shown to be specific and severalfold more sensitive than cultures and clinically useful. Therefore, PCR may be considered the gold standard to establish the etiology of infectious endophthalmitis. As there is no statistically significant difference in the results of PCR on AH and VF, PCR on AH could be the method of choice considering safety and simplicity of the procedure of its collection.
MeSH term(s)	Aqueous Humor/microbiology ; Bacteria/genetics ; Bacteria/isolation & purification ; Cataract Extraction ; DNA, Bacterial/analysis ; DNA, Fungal/analysis ; Endophthalmitis/diagnosis ; Endophthalmitis/microbiology ; Eye Infections, Bacterial/diagnosis ; Eye Infections, Bacterial/microbiology ; Eye Infections, Fungal/diagnosis ; Eye Infections, Fungal/microbiology ; Fungi/genetics ; Fungi/isolation & purification ; Humans ; Polymerase Chain Reaction/methods ; Postoperative Complications ; Vitreous Body/microbiology
Chemical Substances	DNA, Bacterial ; DNA, Fungal
Language	English
Publishing date	2009-09-03
Publishing country	United States
Document type	Journal Article
ZDB-ID	1089461-5
ISSN	1724-6016 ; 1120-6721
ISSN (online)	1724-6016
ISSN	1120-6721
DOI	10.1177/112067210901900520
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Zs.A 3342: Show issues

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Article ; Online: Detection of novel and reported mutations in the rpoB, katG and inhA genes in multidrug-resistant tuberculosis isolates: A hospital-based study.

Ramasubban, Gayathri / Therese, Kulandai Lily / Lakshmipathy, Dhanurekha / Sridhar, R / Meenakshi, N / Madhavan, Hajib N

Journal of global antimicrobial resistance

2015 Volume 3, Issue 1, Page(s) 1–4

Abstract: The objective of this study was to detect mutations associated with isoniazid (INH) and rifampicin (RIF) resistance in Mycobacterium tuberculosis isolates from newly diagnosed and previously treated tuberculosis patients using a PCR-based DNA sequencing ... ...

Abstract	The objective of this study was to detect mutations associated with isoniazid (INH) and rifampicin (RIF) resistance in Mycobacterium tuberculosis isolates from newly diagnosed and previously treated tuberculosis patients using a PCR-based DNA sequencing technique. Phenotypic drug susceptibility testing was performed using a BACTEC™ MicroMGIT Culture System in 354 M. tuberculosis isolates. Among the 354 isolates, 18 were multidrug-resistant tuberculosis (MDR-TB). PCR-based DNA sequencing was performed targeting the rpoB gene for RIF and the whole of the katG gene and the promoter and coding region of the inhA gene for INH. Results were analysed using MultAlin analysis to identify the presence of polymorphisms or mutations by comparing with already available GenBank sequences. Only 37.5% of RIF-resistant isolates showed the presence of the most commonly reported mutation (Ser531Leu). The most commonly reported mutation (Ser531Leu) was detected in six MDR-TB isolates. The frequency of mutations associated with INH resistance was 31.5% (17/54) and 29.6% (16/54) for katG and inhA, respectively. Comparing the relative distribution of mutations in the two target loci revealed that 12 isolates (22.2%) had a mutation in both katG and inhA. Apart from previously reported mutations in the katG gene, there were three novel deletion and six novel substitution mutations. As reported in previous studies, Ser531Leu was the most common mutation detected in RIF-resistant isolates. The genetic mechanism of INH resistance in M. tuberculosis is highly complex involving several genes, and much remains to be explored to achieve a better understanding of this complex mechanism.
Language	English
Publishing date	2015-03
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	2710046-7
ISSN	2213-7173 ; 2213-7165
ISSN (online)	2213-7173
ISSN	2213-7165
DOI	10.1016/j.jgar.2014.10.002
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Article: Virulence genome analysis of Pseudomonas aeruginosa VRFPA10 recovered from patient with scleritis

Murugan, Nandagopal / Malathi, Jambulingam / Umashankar, Vetrivel / Madhavan, Hajib Narahari Rao

Genomics Data. 2016,

2016

Abstract: ... N, 80° 16′′ 13.818′′ E) ...

Abstract	Infectious keratitis is a major cause of blindness, next to cataract and majority of cases are mainly caused by gram negative bacterium Pseudomonas aeruginosa (P. aeruginosa). In this study, we investigated a P. aeruginosa VRFPA10 genomewhich exhibited susceptibility to commonly used drugs in vitro but the patient had poor prognosis due to its hyper virulent nature. Genomic analysis of VRFPA10 deciphered multiple virulence factors and P.aeruginosa Genomic Islands (PAGIs) VRFPA10 genome which correlated with hyper virulence nature of the organism. The genome sequence has been deposited in DDBJ/EMBL/GenBank under the accession numbers LFMZ01000001-LFMZ01000044.Image 1SpecificationsOrganism/cell line/tissuePseudomonas aeruginosaStrainVRFPA10SexN/ASequence or Array typeION PGMData formatAnalyzedExperimental factorsGenomic DNA extracted from pure bacterial culture isolated from the Human scleral scrapingExperimental featuresDraft genome sequence of P. aeruginosa VRFPA02, assembly and annotationConsentApprovedSample Source LocationVision Research Foundation, Sankara Nethralaya Eye Hospital Chennai, Tamil Nadu, India (13° 5′′ 0.1212′′ N, 80° 16′′ 13.818′′ E)
Keywords	Gram-negative bacteria ; Pseudomonas aeruginosa ; blindness ; cataract ; drugs ; genomic islands ; genomics ; keratitis ; nucleotide sequences ; patients ; prognosis ; sequence analysis ; virulence
Language	English
Publishing place	Elsevier Inc.
Document type	Article
Note	Pre-press version
ZDB-ID	2751131-5
ISSN	2213-5960
ISSN	2213-5960
DOI	10.1016/j.gdata.2017.02.007
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Article ; Online: Genotypic Detection of Epstein Barr Virus in Pediatric Transplant Recipients From India.

Janani, Madhuravasal Krishnan / Malathi, Jambulingam / Rela, Mohamed / Farouk, Mohammed / Padmapriya, J / Madhavan, Hajib N

Indian pediatrics

2015 Volume 52, Issue 11, Page(s) 946–950

Abstract: Objective: To determine the rate of occurrence and genotypes of Epstein-Barr Virus (EBV) among pediatric renal and liver transplants recipients.: Design: Observational study.: Setting: Vision Research Foundation referral center and Institute of ... ...

Abstract	Objective: To determine the rate of occurrence and genotypes of Epstein-Barr Virus (EBV) among pediatric renal and liver transplants recipients. Design: Observational study. Setting: Vision Research Foundation referral center and Institute of Liver Disease and Transplantation, Chennai, India. Participants: 70 pediatric solid organ transplant recipients and 60 voluntary healthy donors. Methods: Polymerase chain reaction (PCR) for detection and genotyping of EBV were carried out using genes targeting Viral capsid antigen, Nuclear antigen 1, 2 and 3, followed by real time PCR for viral load determination and further confirmed by phylogenetic analysis. Results: EBV was detected in 35 (51.4%) samples (32 liver and 4 renal transplants) with high viral load. Type A was detected in 33 samples, Type B in 2 liver transplant patients, and co-infection in one liver transplant patient who developed Post-transplant Lymphoproliferative Disorder (PTLD). Real time PCR results correlated with conventional PCR. The mean viral load for patients who did not develop PTLD was 50,424 copies/mL. Overall EBV load in patient with PTLD ranged from 1,40,392 copies/mL prior to PTLD diagnosis to 62,124 copies /mL post treatment. Conclusion: EBV infection is the high risk factor for PTLD after liver transplantation. PCR targeting of EBV can be applied to diagnose EBV infections and monitor treatment for EBV in pediatric solid organ transplant recipients.
MeSH term(s)	Adolescent ; Adult ; Epstein-Barr Virus Infections/etiology ; Epstein-Barr Virus Infections/transmission ; Epstein-Barr Virus Infections/virology ; Herpesvirus 4, Human/genetics ; Humans ; India/epidemiology ; Kidney Transplantation/adverse effects ; Liver Transplantation/adverse effects ; Polymerase Chain Reaction ; Transplant Recipients/statistics & numerical data ; Transplants/virology ; Young Adult
Language	English
Publishing date	2015-11-17
Publishing country	India
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	402594-5
ISSN	0974-7559 ; 0019-6061
ISSN (online)	0974-7559
ISSN	0019-6061
DOI	10.1007/s13312-015-0750-7
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Zs.A 859: Show issues

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Article: A study on the characterization of Propionibacterium acnes isolated from ocular clinical specimens.

Sowmiya, Murali / Malathi, Jambulingam / Swarnali, Sen / Priya, Jeyavel Padma / Therese, Kulandai Lily / Madhavan, Hajib N

The Indian journal of medical research

2015 Volume 142, Issue 4, Page(s) 438–449

Abstract: Background & objectives: There are only a few reports available on characterization of Propionibacterium acnes isolated from various ocular clinical specimens. We undertook this study to evaluate the role of P. acnes in ocular infections and biofilm ... ...

Abstract	Background & objectives: There are only a few reports available on characterization of Propionibacterium acnes isolated from various ocular clinical specimens. We undertook this study to evaluate the role of P. acnes in ocular infections and biofilm production, and also do the phylogenetic analysis of the bacilli. Methods: One hundred isolates of P. acnes collected prospectively from ocular clinical specimens at a tertiary care eye hospital between January 2010 and December 2011, were studied for their association with various ocular disease conditions. The isolates were also subjected to genotyping and phylogenetic analysis, and were also tested for their ability to produce biofilms. Results: Among preoperative conjunctival swabs, P. acnes was a probably significant pathogen in one case; a possibly significant pathogen in two cases. In other clinical conditions, 13 per cent isolates were probably significant pathogens and 38 per cent as possibly significant pathogens. The analysis of 16S rRNA gene revealed four different phylogenies whereas analysis of recA gene showed two phylogenies confirming that recA gene was more reliable than 16S rRNA with less sequence variation. Results of polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) had 100 per cent concordance with phylogenetic results. No association was seen between P. acnes subtypes and biofilm production. Interpretation & conclusions: RecA gene phylogenetic studies revealed two different phylogenies. RFLP technique was found to be cost-effective with high sensitivity and specificity in phylogenetic analysis. No association between P. acnes subtypes and pathogenetic ability was observed. Biofilm producing isolates showed increased antibiotic resistance compared with non-biofilm producing isolates.
MeSH term(s)	Biofilms ; DNA, Bacterial/genetics ; DNA, Bacterial/isolation & purification ; Drug Resistance, Microbial/genetics ; Eye/microbiology ; Eye/pathology ; Eye Diseases/genetics ; Eye Diseases/microbiology ; Eye Diseases/pathology ; Genotype ; Humans ; Phylogeny ; Polymorphism, Restriction Fragment Length ; Propionibacterium acnes/isolation & purification ; Propionibacterium acnes/pathogenicity ; RNA, Ribosomal, 16S/genetics ; Rec A Recombinases/genetics
Chemical Substances	DNA, Bacterial ; RNA, Ribosomal, 16S ; Rec A Recombinases (EC 2.7.7.-)
Language	English
Publishing date	2015-10
Publishing country	India
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	390883-5
ISSN	0971-5916 ; 0019-5340
ISSN	0971-5916 ; 0019-5340
DOI	10.4103/0971-5916.169209
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Article: Whole-Genome Sequencing of Streptomycin-Resistant Mycobacterium tuberculosis Isolate VRFCWCF MRTB 180 Reveals Novel and Potential Mutations for Resistance.

Ramasubban, Gayathri / Lakshmipathy, Dhanurekha / Vetrivel, Umashankar / Kulandai, Lily Therese / Madhavan, Hajib Narahari / Sridhar, R / Meenakshi, N

Genome announcements

2014 Volume 2, Issue 5

Abstract: We announce the draft genome sequence of a streptomycin monoresistant Mycobacterium tuberculosis strain (VRFCWCF MRTB 180) isolated from sputum of a clinically suspected tuberculosis patient. ...

Abstract	We announce the draft genome sequence of a streptomycin monoresistant Mycobacterium tuberculosis strain (VRFCWCF MRTB 180) isolated from sputum of a clinically suspected tuberculosis patient.
Language	English
Publishing date	2014-09-11
Publishing country	United States
Document type	Journal Article
ZDB-ID	2704277-7
ISSN	2169-8287
ISSN	2169-8287
DOI	10.1128/genomeA.00919-14
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Article: Whole Genome Sequence of Polyresistant Mycobacterium tuberculosis CWCFVRF PRTB 19 Sputum Isolate from Chennai, India, Closely Clustering with East African Indian 5 Genogroup.

Lakshmipathy, Dhanurekha / Vetrivel, Umashankar / Ramasubban, Gayathri / Kulandai, Lily Therese / Madhavan, Hajib Narahari / Sridhar, R / Meenakshi, N

Genome announcements

2014 Volume 2, Issue 4

Abstract: We announce the draft genome sequence of a polyresistant Mycobacterium tuberculosis strain (CWCFVRF PRTB 19) isolated from the sputum of a clinically suspected tuberculosis patient, and it closely clusters to the East African Indian 5 (EAI5) lineage. ...

Abstract	We announce the draft genome sequence of a polyresistant Mycobacterium tuberculosis strain (CWCFVRF PRTB 19) isolated from the sputum of a clinically suspected tuberculosis patient, and it closely clusters to the East African Indian 5 (EAI5) lineage.
Language	English
Publishing date	2014-07-17
Publishing country	United States
Document type	Journal Article
ZDB-ID	2704277-7
ISSN	2169-8287
ISSN	2169-8287
DOI	10.1128/genomeA.00702-14
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