Development of a semi-automated colorimetric assay for screening anti-leishmanial agents.


:MTS or {3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl}-2H-tetrazolium, inner salt) is converted into soluble formazan by mitochondrial dehydrogenase of viable cells, thus serving as an indicator of cell viability. Accordingly, a MTS-based assay was developed to evaluate anti-leishmanial activity in Leishmania promastigotes from strains responsible for visceral, cutaneous or mucocutaneous leishmaniasis. The assay was initially optimized for the appropriate wavelength (490 nm), culture medium (M-199), incubation time (3 h) and temperature (37 degrees C). Increasing absorbance with increasing cell density confirmed linearity of the assay that was maintained up to 2.5 x 10(6) cells/200 microl. The growth kinetics of six L. donovani strains and six non-L. donovani strains consistently indicated higher absorbances in the L. donovani strains highlighting the importance of strain-specific customization of the MTS assay. The IC(50) values (i.e., the concentration at which 50% of growth was inhibited) of amphotericin B, miltefosine and pentamidine isethionate obtained by the MTS assay corroborated with previously published data. Taken together, the MTS assay thus permits a simple, reproducible and reliable semi-automated method for evaluating cell viability, effective for drug-screening and growth kinetic studies.


J Microbiol Methods


Ganguly S,Bandyopadhyay S,Sarkar A,Chatterjee M




Has Abstract


2006-07-01 00:00:00














  • Amplified ribosomal DNA restriction analysis for the characterization of Azotobacteraceae: a contribution to the study of these free-living nitrogen-fixing bacteria.

    abstract::A 16S rRNA gene-based fingerprinting method was developed for the identification of Azotobacteraceae and tested onto 48 soil isolates and 28 reference strains belonging to the free-living nitrogen-fixing bacterial group and to the most common species found in soil samples. According to this method, the 16S rRNA gene w...

    journal_title:Journal of microbiological methods

    pub_type: 杂志文章


    authors: Aquilanti L,Mannazzu I,Papa R,Cavalca L,Clementi F

    更新日期:2004-05-01 00:00:00

  • Use of both 16S rRNA and engineered functional genes with real-time PCR to quantify an engineered, PCB-degrading Rhodococcus in soil.

    abstract::A real-time PCR (RTm-PCR) assay using fluorescently labeled oligonucleotides (TaqMan probes) was used to detect and quantify the recombinant Rhodococcus sp. strain RHA1(fcb) in soil. One primer and probe set targeted a hypervariable region of the 16S rRNA gene unique to strain RHA1(fcb) and its phylogenetic relatives,...

    journal_title:Journal of microbiological methods

    pub_type: 杂志文章


    authors: Rodrigues JL,Aiello MR,Urbance JW,Tsoi TV,Tiedje JM

    更新日期:2002-10-01 00:00:00

  • Microextraction and Gas Chromatography/Mass Spectrometry for improved analysis of geosmin and other fungal "off" volatiles in grape juice.

    abstract::Geosmin is a volatile fungal metabolite with an earthy aroma produced in grape products from rotten grapes. The accumulation of geosmin in grapes is caused by the interaction of Botrytis cinerea and Penicillium expansum. Solid Phase Microextraction (SPME) has great utility for collecting volatile compounds in wine. Ho...

    journal_title:Journal of microbiological methods

    pub_type: 杂志文章


    authors: Morales-Valle H,Silva LC,Paterson RR,Oliveira JM,Venâncio A,Lima N

    更新日期:2010-10-01 00:00:00

  • Improved detection of microbial DNA after bead-beating before DNA isolation.

    abstract::Detection of microbial DNA in clinical samples by real-time PCR requires a universal extraction method with equally efficient lysis of cell walls of all possible microorganisms. We demonstrated that physical disruption by bead-beating with 0.1mm beads in combination with MagNA Pure DNA III extraction enhances microbia...

    journal_title:Journal of microbiological methods

    pub_type: 杂志文章


    authors: de Boer R,Peters R,Gierveld S,Schuurman T,Kooistra-Smid M,Savelkoul P

    更新日期:2010-02-01 00:00:00

  • High density whole genome fingerprinting of methicillin-resistant and -susceptible strains of Staphylococcus aureus in search of phenotype-specific molecular determinants.

    abstract::AFLP is a selective restriction fragment amplification method generating DNA fingerprints for microbial isolates. We present high-throughput AFLP (htAFLP) to characterize molecular markers associated with bacterial phenotypes. Methicillin-resistant and -susceptible isolates of Staphylococcus aureus have been used for ...

    journal_title:Journal of microbiological methods

    pub_type: 杂志文章


    authors: Savelkoul PH,Melles DC,Buffing N,Gorkink R,Simons G,van Belkum A

    更新日期:2007-10-01 00:00:00

  • Multiplex and quantifiable detection of nucleic acid from pathogenic fungi using padlock probes, generic real time PCR and specific suspension array readout.

    abstract::A new concept for multiplex detection and quantification of microbes is here demonstrated on a range of infectious fungal species. Padlock probe methodology in conjunction with qPCR and Luminex technology was used for simultaneous detection of ten fungal species in one single experiment. By combining the multiplexing ...

    journal_title:Journal of microbiological methods

    pub_type: 杂志文章


    authors: Eriksson R,Jobs M,Ekstrand C,Ullberg M,Herrmann B,Landegren U,Nilsson M,Blomberg J

    更新日期:2009-08-01 00:00:00

  • Normalization of test and evaluation of biothreat detection systems: overcoming microbial air content fluctuations by using a standardized reagent bacterial mixture.

    abstract::Test and evaluation of engineered biothreat agent detection systems ("biodetectors") are a challenging task for government agencies and industries involved in biosecurity and biodefense programs. In addition to user friendly features, biodetectors need to perform both highly sensitive and specific detection, and must ...

    journal_title:Journal of microbiological methods

    pub_type: 杂志文章


    authors: Berchebru L,Rameil P,Gaudin JC,Gausson S,Larigauderie G,Pujol C,Morel Y,Ramisse V

    更新日期:2014-10-01 00:00:00

  • Modification of the classical Lancefield assay of group A streptococcal killing to reduce inter-donor variation.

    abstract::The lack of a surrogate-of-immunity assay presents a major barrier to Streptococcus pyogenes research. Modification of the Lancefield assay to include an antibody digestion step reduced inter-donor variation and permitted detection of the anti-streptococcal activity of intravenous immunoglobulin and convalescent serum...

    journal_title:Journal of microbiological methods

    pub_type: 杂志文章


    authors: Reglinski M,Lynskey NN,Sriskandan S

    更新日期:2016-05-01 00:00:00

  • A preliminary evaluation of a new enzyme immunoassay to detect Chlamydia pneumoniae-specific antibodies.

    abstract::New enzyme immunoassays (EIAs) for determination of specific IgG, IgA, and IgM antibody titers to Chlamydia pneumoniae were evaluated independently in three research laboratories. Specificity of the EIAs was enhanced by removing LPS from the chlamydial antigen. The performance of these EIAs was evaluated in comparison...

    journal_title:Journal of microbiological methods

    pub_type: 临床试验,杂志文章,多中心研究


    authors: Ossewaarde JM,Tuuminen T,Boersma WG,Sandström M,Palomäki P,Boman J

    更新日期:2000-12-15 00:00:00

  • Real-time reverse transcription PCR for the quantification of the mntH expression of Salmonella enterica as a function of growth phase and phagosome-like conditions.

    abstract::This article presents an experimental design for measuring the mRNA expression in Salmonella enterica of the mntH gene in phagosome-mimicking conditions. The expression of mntH was quantified by real-time reverse transcription PCR for different S. enterica strains of porcine origin under different biological growth co...

    journal_title:Journal of microbiological methods

    pub_type: 杂志文章


    authors: Botteldoorn N,Van Coillie E,Grijspeerdt K,Werbrouck H,Haesebrouck F,Donné E,D'Haese E,Heyndrickx M,Pasmans F,Herman L

    更新日期:2006-07-01 00:00:00

  • Microscopic quantification of bacterial invasion by a novel antibody-independent staining method.

    abstract::Microscopic discrimination between extracellular and invasive, intracellular bacteria is a valuable technique in microbiology and immunology. We describe a novel fluorescence staining protocol, called FITC-biotin-avidin (FBA) staining, which allows the differentiation between extracellular and intracellular bacteria a...

    journal_title:Journal of microbiological methods

    pub_type: 杂志文章


    authors: Agerer F,Waeckerle S,Hauck CR

    更新日期:2004-10-01 00:00:00

  • Comparative studies on measurement of membrane potential of bacterial cells treated with ZnO nanoparticles by Spectrofluorometry, fluorescence microscopy and flowcytometry.

    abstract::Many methods are developed to assess antimicrobial action of ZnO nanoparticles (NPs). A large number of methods associated with the use of fluorescent probes are developed, including Spectrofluorometry, fluorescence microscopy, and cytometry. In this study, flowcytometry, Spectrofluorometry and fluorescent microscopy ...

    journal_title:Journal of microbiological methods

    pub_type: 杂志文章


    authors: Khater M,Khater SS,Gholap H,Patil R,Kulkarni G

    更新日期:2020-06-01 00:00:00

  • Electroporation as a tool to transfer the plasmid pRL489 in Oscillatoria MKU 277.

    abstract::To establish the gene transfer system in the cyanobacteria, we successfully introduced the shuttle plasmid pRL489 into Oscillatoria MKU 277 by electroporation with transformation frequencies of up to 102 cfu/mug of plasmid DNA. These findings suggest that Oscillatoria MKU 277 can be used as an experimental tool for ge...

    journal_title:Journal of microbiological methods

    pub_type: 杂志文章


    authors: Ravindran CR,Suguna S,Shanmugasundaram S

    更新日期:2006-07-01 00:00:00

  • A new filter-eluting solution that facilitates improved recovery of Cryptosporidium oocysts from water.

    abstract::Cryptosporidium is a zoonotic coccidian parasite associated with diarrhea, and the disinfectant-resistant oocysts are threats to public health even in industrialized countries. In order to make an accurate assessment of the risk to public health, a detection method that has a high recovery rate of oocysts in water is ...

    journal_title:Journal of microbiological methods

    pub_type: 杂志文章


    authors: Inoue M,Rai SK,Oda T,Kimura K,Nakanishi M,Hotta H,Uga S

    更新日期:2003-12-01 00:00:00

  • Use of denaturing gradient gel electrophoresis for analysis of the stool microbiota of hospitalized patients.

    abstract::Denaturing gradient gel electrophoresis (DGGE) of PCR-amplified ribosomal RNA gene amplicons was used to study the stool microbiota of hospitalized patients and to examine the effect of antibiotic therapy. For one patient, 16 anaerobic species identified by random cloning and sequencing of PCR-amplified rRNA genes fro...

    journal_title:Journal of microbiological methods

    pub_type: 杂志文章


    authors: Donskey CJ,Hujer AM,Das SM,Pultz NJ,Bonomo RA,Rice LB

    更新日期:2003-08-01 00:00:00

  • Optimization of electrotransformation (ETF) conditions in lactic acid bacteria (LAB).

    abstract::Lactic acid bacteria (LAB) play important roles in the food industry, animal husbandry and medicine which are closely related to human life. Modern gene engineering technology is the major means to reveal gene functions, study metabolic pathways and metabolomes of strains and improve the properties of strains. However...

    journal_title:Journal of microbiological methods

    pub_type: 杂志文章,评审


    authors: Wang C,Cui Y,Qu X

    更新日期:2020-07-01 00:00:00

  • Use of green fluorescent protein to quantify the growth of Colletotrichum during infection of tobacco.

    abstract::To develop a quantitative assay of fungal growth inside plant tissues, strains of Colletotrichum destructivum and Colletotrichum orbiculare were transformed with a modified green fluorescent protein (GFP) gene fused with a glyceraldehyde-3-phosphate dehydrogenase promoter from Aspergillus nidulans. Transformants expre...

    journal_title:Journal of microbiological methods

    pub_type: 杂志文章


    authors: Chen N,Hsiang T,Goodwin PH

    更新日期:2003-04-01 00:00:00

  • Development of a multiplex PCR assay based on the pilA gene sequences to detect different types of Acidovorax citrulli.

    abstract::Bacterial fruit blotch (BFB) of cucurbits, caused by Acidovorax citrulli, is a major threat to commercial watermelon and melon production worldwide. At present, there are at least two genetically distinct sub-populations (group I and II) of A. citrulli that differ in host preference among cucurbit species and copper s...

    journal_title:Journal of microbiological methods

    pub_type: 杂志文章


    authors: Yang YW,Zhao M,Zhang LQ,Qiao P,Bai X,Zhang XX,Walcott RR,Guan W,Zhao TC

    更新日期:2019-03-01 00:00:00

  • A novel nucleic lateral flow assay for screening of PHA-producing haloarchaea.

    abstract::Polyhydroxyalkanoates (PHAs) are important for biodegradable plastic production, and prokaryotes play a very important role in PHA production. PHA synthase is a key enzyme for the polymerization of PHAs. There are four classes of PHA synthase. The phaC gene is necessary for the production of all classes of PHA synthas...

    journal_title:Journal of microbiological methods

    pub_type: 杂志文章


    authors: Muangsuwan W,Ruangsuj P,Chaichanachaicharn P,Yasawong M

    更新日期:2015-09-01 00:00:00

  • Quantitation of Helicobacter pylori ureC gene and its comparison with different diagnostic techniques and gastric histopathology.

    abstract::Numerous diagnostic assays for Helicobacter pylori detection are available. However, these techniques have their own advantages as well as limitations. Here we tried to develop a real-time quantitative (Q) PCR assay to measure ureC copy number to detect H. pylori, based on the fact that there is only one copy of the u...

    journal_title:Journal of microbiological methods

    pub_type: 杂志文章


    authors: Shukla SK,Prasad KN,Tripathi A,Ghoshal UC,Krishnani N,Nuzhat H

    更新日期:2011-08-01 00:00:00

  • Comparison of culture and biochemical tests with PCR for detection of Brachyspira hyodysenteriae and Brachyspira pilosicoli.

    abstract::Traditional culture and biochemical tests (CBT) were compared with PCR for sensitivity and detection of Brachyspira hyodysenteriae and Brachyspira pilosicoli in seeded faeces and clinical samples from diarrhoeic pigs. A duplex PCR system was developed based on primers detecting the tlyA-gene of B. hyodysenteriae and t...

    journal_title:Journal of microbiological methods

    pub_type: 杂志文章


    authors: Råsbäck T,Fellström C,Gunnarsson A,Aspán A

    更新日期:2006-08-01 00:00:00

  • Evaluation of methodologies including immunofluorescent assay (IFA) and the polymerase chain reaction (PCR) for detection of human pathogenic microsporidia in water.

    abstract::Microsporidia is a term used to describe a group of emerging protozoan pathogens whose environmental occurrence has only recently been documented due to lack of detection methodologies. This study evaluates and describes current methods for detection of microsporidia in water. Standard methods, for the collection and ...

    journal_title:Journal of microbiological methods

    pub_type: 杂志文章


    authors: Dowd SE,Gerba CP,Kamper M,Pepper IL

    更新日期:1999-02-01 00:00:00

  • Metagenomic analysis of bronchoalveolar lavage samples from patients with idiopathic interstitial pneumonia and its antagonic relation with Pneumocystis jirovecii colonization.

    abstract::Idiopathic interstitial pneumonias are interstitial lung diseases of unknown etiology which prognosis is usually fatal. Microbiota associated to bronchoalveolar lavage from 20 patients with negative bacterial cultures was explored by 16S-rDNA PCR-DGGE, showing a clearly negative relation among the presence of P. jirov...

    journal_title:Journal of microbiological methods

    pub_type: 杂志文章


    authors: Friaza V,la Horra Cd,Rodríguez-Domínguez MJ,Martín-Juan J,Cantón R,Calderón EJ,Del Campo R

    更新日期:2010-07-01 00:00:00

  • A strategy to verify the absence of the pgm locus in Yersinia pestis strain candidates for select agent exemption.

    abstract::Yersinia pestis is a Department of Health and Human Services select agent as defined in federal regulations. Certain attenuated strains of Y. pestis, such as the pgm(-) strain, are exempt from these regulations. Herein we describe a strategy to verify the absence of the pgm locus in Y. pestis strains being considered ...

    journal_title:Journal of microbiological methods

    pub_type: 杂志文章


    authors: Jenkins AL,Worsham PL,Welkos SL

    更新日期:2009-06-01 00:00:00

  • Pushing the limits for amplifying BrdU-labeled DNA encoding 16S rRNA: DNA polymerase as the determining factor.

    abstract::Identifying microorganisms that are active under specific conditions in ecosystems is a challenge in microbial ecology. Recently, the bromodeoxyuridine (BrdU) technique was developed to label actively growing cells. BrdU, a thymidine analog, is incorporated into newly synthesized DNA, and the BrdU-labeled DNA is then ...

    journal_title:Journal of microbiological methods

    pub_type: 杂志文章


    authors: Roux-Michollet DD,Schimel JP,Holden PA

    更新日期:2010-12-01 00:00:00

  • Laser interferometric and cultivation methods for measurement of colistin/ampicilin and saponin interactions with smooth and rough of Proteus mirabilis lipopolysaccharides and cells.

    abstract::Laser interferometry is commonly used in permeability studies of soluble substances. In this study a modification that allowed testing partially insoluble mixtures is presented. The modification relies on the measurement of diffusion from 1% agarose gel. As a model for this study, two Proteus mirabilis strains were us...

    journal_title:Journal of microbiological methods

    pub_type: 杂志文章


    authors: Arabski M,Wasik S,Dworecki K,Kaca W

    更新日期:2009-05-01 00:00:00

  • Rapid detection of "highly virulent" Group B Streptococcus ST-17 and emerging ST-1 clones by MALDI-TOF mass spectrometry.

    abstract::MALDI-TOF MS identified a 6250-Da protein specific to Sequence Type-1 (ST-1) strains and a 7625-Da protein specific to ST-17 strains when used for identification of Group B streptococci. The strains of these STs are major causes of meningitis and late-onset-disease in neonates. This rapid method of identification coul...

    journal_title:Journal of microbiological methods

    pub_type: 杂志文章


    authors: Lartigue MF,Kostrzewa M,Salloum M,Haguenoer E,Héry-Arnaud G,Domelier AS,Stumpf S,Quentin R

    更新日期:2011-08-01 00:00:00

  • Development of an oriC vector for use in Mycoplasma synoviae.

    abstract::Mycoplasma synoviae, an important poultry pathogen, belonging to the class Mollicutes, causes airsacculitis, synovitis, decreased egg production and produces significant economic losses. Efforts to determine M. synoviae virulence factors and their role in pathogenicity require suitable tools for genetic manipulation o...

    journal_title:Journal of microbiological methods

    pub_type: 杂志文章


    authors: Shahid MA,Marenda MS,Markham PF,Noormohammadi AH

    更新日期:2014-08-01 00:00:00

  • Development of a novel plasmid vector pTIO-1 adapted for electrotransformation of Porphyromonas gingivalis.

    abstract::We report here the construction of a plasmid vector designed for the efficient electrotransformation of the periodontal pathogen Porphyromonas gingivalis. The novel Escherichia coli-Bacteroides/P. gingivalis shuttle vector, designated pTIO-1, is based on the 11.0-kb E. coli-Bacteroides conjugative shuttle vector, pVAL...

    journal_title:Journal of microbiological methods

    pub_type: 杂志文章


    authors: Tagawa J,Inoue T,Naito M,Sato K,Kuwahara T,Nakayama M,Nakayama K,Yamashiro T,Ohara N

    更新日期:2014-10-01 00:00:00

  • Isolation of Harveyi clade Vibrio spp. collected in aquaculture farms: How can the identification issue be addressed?

    abstract::Aquaculture is a fast growing industry with its development hampered by bacterial diseases. Vibriosis caused by Harveyi clade strains is known for causing heavy loss especially in shrimp aquaculture farms. For farm treatment and pathogen spread management, veterinarians and researchers need reliable bacterial identifi...

    journal_title:Journal of microbiological methods

    pub_type: 杂志文章


    authors: Culot A,Grosset N,Bruey Q,Auzou M,Giard JC,Favard B,Wakatsuki A,Baron S,Frouel S,Techer C,Gautier M

    更新日期:2021-01-01 00:00:00