Bacteriophage tail-spike protein derivitized microresonator arrays for specific detection of pathogenic bacteria

2013 ◽  
Vol 181 ◽  
pp. 410-416 ◽  
Author(s):  
Somayyeh Poshtiban ◽  
Amit Singh ◽  
Glen Fitzpatrick ◽  
Stephane Evoy
Keyword(s):  
Pathogenic Bacteria ◽  
Spike Protein ◽  
Specific Detection ◽  
Tail Spike

Fate and transport of faecal contamination microbial indicators, pathogenic protozoa and Campylobacter in the artificially recharged fractured aquifer of Salento, Italy

2008 ◽  
Vol 57 (6) ◽  
pp. 849-856 ◽  
Author(s):  
R. La Mantia ◽  
C. Masciopinto ◽  
C. Levantesi ◽  
V. Tandoi
Keyword(s):  
Pathogenic Bacteria ◽  
Fate And Transport ◽  
Fractured Aquifer ◽  
Specific Detection ◽  
Microbial Indicators ◽  
Faecal Contamination ◽  
Time Required ◽  
Indicators Of Faecal Contamination ◽  
Campylobacter Spp

The study investigates the fate and transport of microorganisms introduced by artificial groundwater recharge at the Nardò fractured aquifer in Salento, Italy. Microbial indicators of faecal contamination, parasitic protozoa (Giardia and Cryptosporidium) and pathogenic bacteria (Campylobacter spp.), were monitored into injected water and groundwater to test the efficiency of the “natural disinfection” into the fractured aquifer. A remarkable decrease of microbial indicators and pathogens was observed suggesting that pathogens removal or inactivation may be possible during water flow in fractured aquifer. The recently described PNA probe CJE195 (Lehtola et al. 2005) was utilised for the rapid and specific detection of Campylobacter spp. by fluorescence in situ hybridization (FISH) after enrichment. FISH results were consistent with those of traditional cultural method (ISO 17995) applied in parallel: time required for Campylobacter identification was reduced of 4 days.

scholarly journals A Multiplex-PCR Method for Strain Identification and Detailed Phylogenetic Analysis of AY-Group Phytoplasmas

Plant Disease ◽  
2014 ◽  
Vol 98 (3) ◽  
pp. 299-305 ◽  
Author(s):  
Shigeyuki Kakizawa ◽  
Yoichi Kamagata
Keyword(s):  
Phylogenetic Analysis ◽  
Multiplex Pcr ◽  
16S Rdna ◽  
Pathogenic Bacteria ◽  
Target Genes ◽  
Direct Sequencing ◽  
Pcr Amplification ◽  
Single Copy ◽  
Strain Identification ◽  
Specific Detection

Phytoplasmas are plant pathogenic bacteria that cause devastating losses in the yield of diverse crops worldwide. Specific detection and strain identification of phytoplasmas is important to prevent the spread of phytoplasma-induced diseases. Hence, methods to rapidly detect these organisms are important for pest control. Polymerase chain reaction (PCR) methods using phytoplasma-specific primers are widely used to detect phytoplasmas from infected plants and insects because they are highly sensitive, easily handled, and have a variety of analytical secondary applications. The phytoplasma 16S rDNA was widely used as a target of the PCR detection method; however, further target genes and more rapid methods have been required for more specific detection of phytoplasmas. Here, we developed a multiplex-PCR system to amplify several phytoplasma genes. We designed 36 primers, based on the genome sequence of ‘Candidatus Phytoplasma asteris’, to amplify 18 single-copy genes covering wide regions of the phytoplasma genome. Nine genes could be simultaneously amplified in a single PCR. This multiplex-PCR was applied to DNAs from 10 phytoplasma strains belonging to the AY-group, and different amplification patterns were obtained between strains, suggesting that this method would allow us to differentiate phytoplasmas at the strain level. Direct sequencing was also possible after the multiplex-PCR amplification by a modified sequencing method. Detailed phylogenetic analysis was performed using concatenated sequences, and evolutionary relationships among four Japanese isolates were revealed, where these strains could not be distinguished by their 16S rDNA. Thus, this multiplex-PCR system is useful for rapid strain identification and detailed phylogenetic analysis of phytoplasmas.

scholarly journals Engineered Reporter Phages for Rapid Bioluminescence-Based Detection and Differentiation of Viable Listeria Cells

2020 ◽  
Vol 86 (11) ◽  
Author(s):  
Susanne Meile ◽  
Anne Sarbach ◽  
Jiemin Du ◽  
Markus Schuppler ◽  
Carmen Saez ◽  
...  
Keyword(s):  
Food Production ◽  
Pathogenic Bacteria ◽  
False Negative ◽  
Host Cells ◽  
Target Cells ◽  
Broad Host Range ◽  
Specific Detection ◽  
Production Chain ◽  
Content Type ◽  
Culture Dependent

ABSTRACT The pathogen Listeria monocytogenes causes listeriosis, a severe foodborne disease associated with high mortality. Rapid and sensitive methods are required for specific detection of this pathogen during food production. Bioluminescence-based reporter bacteriophages are genetically engineered viruses that infect their host cells with high specificity and transduce a heterologous luciferase gene whose activity can be detected with high sensitivity to indicate the presence of viable target cells. Here, we use synthetic biology for de novo genome assembly and activation as well as CRISPR-Cas-assisted phage engineering to construct a set of reporter phages for the detection and differentiation of viable Listeria cells. Based on a single phage backbone, we compare the performance of four reporter phages that encode different crustacean, cnidarian, and bacterial luciferases. From this panel of reporter proteins, nanoluciferase (NLuc) was identified as a superior enzyme and was subsequently introduced into the genomes of a broad host range phage (A511) and two serovar 1/2- and serovar 4b/6a-specific Listeria phages (A006 and A500, respectively). The broad-range NLuc-based phage A511::nlucCPS detects one CFU of L. monocytogenes in 25 g of artificially contaminated milk, cold cuts, and lettuce within less than 24 h. In addition, this reporter phage successfully detected Listeria spp. in potentially contaminated natural food samples without producing false-positive or false-negative results. Finally, A006::nluc and A500::nluc enable serovar-specific Listeria diagnostics. In conclusion, these NLuc-based reporter phages enable rapid, ultrasensitive detection and differentiation of viable Listeria cells using a simple protocol that is 72 h faster than culture-dependent approaches. IMPORTANCE Culture-dependent methods are the gold standard for sensitive and specific detection of pathogenic bacteria within the food production chain. In contrast to molecular approaches, these methods detect viable cells, which is a key advantage for foods generated from heat-inactivated source material. However, culture-based diagnostics are typically much slower than molecular or proteomic strategies. Reporter phage assays combine the best of both worlds and allow for near online assessment of microbial safety because phage replication is extremely fast, highly target specific, and restricted to metabolically active host cells. In addition, reporter phage assays are inexpensive and do not require highly trained personnel, facilitating their on-site implementation. The reporter phages presented in this study not only allow for rapid detection but also enable an early estimation of the potential virulence of Listeria isolates from food production and processing sites.

scholarly journals Specific serology for emerging human coronaviruses by protein microarray

2013 ◽  
Vol 18 (14) ◽  
Author(s):  
C Reusken ◽  
H Mou ◽  
G J Godeke ◽  
L van der Hoek ◽  
B Meyer ◽  
...  
Keyword(s):  
Receptor Binding ◽  
Protein Microarray ◽  
Spike Protein ◽  
Microarray Technology ◽  
Igg Antibodies ◽  
Specific Detection ◽  
Human Coronavirus ◽  
Serological Assay ◽  
Human Coronaviruses ◽  
Binding Subunit

We present a serological assay for the specific detection of IgM and IgG antibodies against the emerging human coronavirus hCoV-EMC and the SARS-CoV based on protein microarray technology. The assay uses the S1 receptor-binding subunit of the spike protein of hCoV-EMC and SARS-CoV as antigens. The assay has been validated extensively using putative cross-reacting sera of patient cohorts exposed to the four common hCoVs and sera from convalescent patients infected with hCoV-EMC or SARS-CoV.

scholarly journals Morphologically Different Pectobacterium brasiliense Bacteriophages PP99 and PP101: Deacetylation of O-Polysaccharide by the Tail Spike Protein of Phage PP99 Accompanies the Infection

2020 ◽  
Vol 10 ◽  
Author(s):  
Anna A. Lukianova ◽  
Mikhail M. Shneider ◽  
Peter V. Evseev ◽  
Anna M. Shpirt ◽  
Eugenia N. Bugaeva ◽  
...  
Keyword(s):  
Spike Protein ◽  
Tail Spike

scholarly journals Rapid and Specific Detection of Burkholderia glumae in Rice Seed by Real-Time Bio-PCR Using Species-Specific Primers Based on an rhs Family Gene

Plant Disease ◽  
2012 ◽  
Vol 96 (4) ◽  
pp. 577-580 ◽  
Author(s):  
Byoung Kyu Kim ◽  
Min Seok Cho ◽  
Myeong Ho Kim ◽  
Hyeon Jin Choi ◽  
Man Jung Kang ◽  
...  
Keyword(s):  
Real Time ◽  
Pathogenic Bacteria ◽  
Quantitative Polymerase Chain Reaction ◽  
Pcr Primers ◽  
Effective Control ◽  
Sequence Information ◽  
Rice Seed ◽  
Specific Detection ◽  
Burkholderia Glumae ◽  
Family Gene

In this study, we developed a reliable, quick, and accurate quantitative polymerase chain reaction (qPCR) assay to detect grain rot caused by Burkholderia glumae in rice seed. The control of bacterial grain rot is difficult, and the only practical methods for disease management rely on the use of pathogen-free seed, appropriate culture practices, and resistant cultivars. Therefore, the specific detection of this pathogen in seed is essential for effective control of the disease. However, other Burkholderia spp. are also detected by currently available molecular and serological methods. In this study, we exploited the available genome sequence information in public databases to develop specific PCR primers for accurate diagnosis of B. glumae. An SYBR Green real-time PCR primer set was designed based on the rhs family gene (YD repeat protein) of B. glumae BGR1 because these genes are structurally diverse. The specificity of the primers was evaluated using purified DNA from 5 isolates of B. glumae, 6 different species of Burkholderia, and 18 other reference pathogenic bacteria. The assay was able to detect at least one genome equivalent of cloned amplified target DNA using purified DNA or 1 CFU per reaction when using calibrated cell suspension. This method is rapid and reliable and has great potential for analyzing large numbers of samples.

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