Multiplex PCR using YeaD and 16S rRNA gene to identify major pathogens in vibriosis of Litopenaeus vannamei

The use of local probiotics in the culture of aquatic organisms is increasing with the demand for more environmental-friendly aquaculture practices. The local bacterium isolate considered as a probiotic was added into the water of whiteleg shrimp (Litopenaeus vannamei) culture in a field trial. Four rectangular plastic ponds (ca. 20 m x 30 m per pond) were used for 100 days experimentation for six consecutive crops in two years experiment. Survival, harvest size, feed conversion ratio (FCR) and Vibrio bacterial count was compared with those of shrimp receiving and none of local isolate. Identification based on 16S rRNA gene sequence shown those isolate was Bacillus pumilus strain DURCK14 with 99% homology. Water shrimp pond added a local isolate had significantly higher survival at about 10.0% to 11.7% than shrimp without added the isolate (p<0.05), and better FCR, but no significant different in shrimp harvest size. Vibrio bacterial was undetected by total plate count. Moreover, it shown better projected yields on an annual basis (three crops per year).

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Rapid identification of Saccharomonospora strains by multiplex PCR using species-specific primers within the 16S rRNA gene

Journal of Microbiological Methods ◽

10.1016/0167-7012(96)00933-5 ◽

1996 ◽

Vol 27 (1) ◽

pp. 89-95 ◽

Cited By ~ 7

Author(s):

Jung-Hoon Yoon ◽

Sung Taik Lee ◽

Yong Kook Shin ◽

Sam-Bong Kim ◽

Hong-Joong Kim ◽

...

Keyword(s):

16S Rrna ◽

16S Rrna Gene ◽

Multiplex Pcr ◽

Rapid Identification ◽

Rrna Gene ◽

Specific Primers ◽

Species Specific ◽

The 16S Rrna Gene

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A novel multiplex PCR/RFLP assay for the identification of Streptococcus bovis/Streptococcus equinus complex members from dairy microbial communities based on the 16S rRNA gene

FEMS Microbiology Letters ◽

10.1111/j.1574-6968.2011.02443.x ◽

2011 ◽

Vol 326 (2) ◽

pp. 144-150 ◽

Cited By ~ 19

Author(s):

Christoph Jans ◽

Christophe Lacroix ◽

Leo Meile

Keyword(s):

16S Rrna ◽

16S Rrna Gene ◽

Microbial Communities ◽

Multiplex Pcr ◽

Rrna Gene ◽

Streptococcus Bovis ◽

Pcr Rflp ◽

Rflp Assay ◽

The 16S Rrna Gene ◽

Streptococcus Equinus

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Development of a simple, rapid multiplex PCR tool kit by using 16S rRNA gene for the identification of faecal and non-faecal coli forms in the drinking waters

Water Science & Technology Water Supply ◽

10.2166/ws.2021.081 ◽

2021 ◽

Author(s):

A. Shiva Shanker ◽

N. Rajesh ◽

Pavan Kumar Pindi

Keyword(s):

Drinking Water ◽

16S Rrna ◽

16S Rrna Gene ◽

Multiplex Pcr ◽

Faecal Coliform ◽

Rrna Gene ◽

Faecal Coliforms ◽

Variable Regions ◽

Total Coliforms ◽

Multiplex Pcr Assay

Abstract A multiplex method for the detection of faecal and non-faecal coliforms in drinking water was developed using three primers from the V2, V3 and V9 variable regions of 16S rRNA gene. 194F, 474F and 1436R are the three primers designed for specific amplification of V2, V3, V9 hyper variable regions of 16S rRNA gene. Multiplex PCR allowed for differentiation of the total coliform from faecal coliform by specific amplicons: 1,285 bp of amplicon is specific for 6 non-faecal coliform genera and 1,009 bp of amplicon is specific for faecal coliform ie. E. coli. If the drinking water was contaminated with both faecal and non-faecal coliforms then two amplicons of 1,285 bp and 1,009 bp by combination of three primers are observed. The multiplex PCR assay based on 16S rRNA gene should be a beneficial tool kit for the rapid identification of the total coliforms in the large number of water samples compared with traditional methods. Results can be acquired within 3 hrs of time as compared with classic method of MPN (3–4 days). This assay will be useful in diversification and detection of seven genera of total coliforms by using variable regions of 16S rRNA.

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Multiplex PCR with 16S rRNA Gene-Targeted Primers of Bifidobacterium spp. To Identify Sources of Fecal Pollution

Applied and Environmental Microbiology ◽

10.1128/aem.00536-07 ◽

2007 ◽

Vol 73 (9) ◽

pp. 3122-3122 ◽

Cited By ~ 1

Author(s):

X. Bonjoch ◽

E. Ballesté ◽

A. R. Blanch

Keyword(s):

16S Rrna ◽

16S Rrna Gene ◽

Multiplex Pcr ◽

Fecal Pollution ◽

Rrna Gene

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Development of a rapid identification method forAeromonasspecies by multiplex-PCR

Canadian Journal of Microbiology ◽

10.1139/w05-089 ◽

2005 ◽

Vol 51 (11) ◽

pp. 957-966 ◽

Cited By ~ 22

Author(s):

Keya Sen

Keyword(s):

16S Rrna ◽

16S Rrna Gene ◽

Multiplex Pcr ◽

Rapid Identification ◽

Complete Agreement ◽

Aeromonas Species ◽

Rrna Gene ◽

Primer Sets ◽

Species Specific ◽

The 16S Rrna Gene

Existing biochemical methods cannot distinguish among some species of Aeromonads, while genetic methods are labor intensive. In this study, primers were developed to three genes of Aeromonas: lipase, elastase, and DNA gyraseB. In addition, six previously described primer sets, five corresponding to species-specific signature regions of the 16S rRNA gene from A. veronii, A. popoffii, A. caviae, A. jandaei, and A. schubertii, respectively, and one corresponding to A. hydrophila specific lipase (hydrolipase), were chosen. The primer sets were combined in a series of multiplex-PCR (mPCR) assays against 38 previously characterized strains. Following PCR, each species was distinguished by the production of a unique combination of amplicons. When the assays were tested using 63 drinking water isolates, there was complete agreement in the species identification (ID) for 59 isolates, with ID established by biochemical assays. Sequencing the gyrB and the 16S rRNA gene from the remaining four strains established that the ID obtained by mPCR was correct for three strains. For only one strain, no consensus ID could be obtained. A rapid and reliable method for identification of different Aeromonas species is proposed that does not require restriction enzyme digestions, thus simplifying and speeding up the process.Key words: Aeromonas, multiplex-PCR, identification.

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