Application of a real-time qPCR method to measure the methanogen concentration during anaerobic digestion as an indicator of biogas production capacity

ABSTRACT A real-time quantitative PCR method was developed for the detection and enumeration of Mycobacterium spp. from environmental samples and was compared to two other methods already described. The results showed that our method, targeting 16S rRNA, was more specific than the two previously published real-time quantitative PCR methods targeting another 16S rRNA locus and the hsp65 gene (100% versus 44% and 91%, respectively).

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SYBR Green real-time qPCR method: Diagnose drowning more rapidly and accurately

Forensic Science International ◽

10.1016/j.forsciint.2021.110720 ◽

2021 ◽

Vol 321 ◽

pp. 110720

Author(s):

Zhonghao Yu ◽

Quyi Xu ◽

Cheng Xiao ◽

Huan Li ◽

Weibin Wu ◽

...

Keyword(s):

Real Time ◽

Sybr Green ◽

Qpcr Method ◽

Real Time Qpcr

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IDENTIFIKASI ZIGOSITAS IKAN LELE (Clarias gariepinus) TRANSGENIK F-2 YANG MEMBAWA GEN HORMON (PhGH) DENGAN MENGGUNAKAN METODE REALTIME-qPCR

Jurnal Riset Akuakultur ◽

10.15578/jra.11.1.2016.39-46 ◽

2016 ◽

Vol 11 (1) ◽

pp. 39

Author(s):

Huria Marnis ◽

Bambang Iswanto ◽

Romy Suprapto ◽

Imron Imron ◽

Raden Roro Sri Pudji Sinarni Dewi

Keyword(s):

Real Time ◽

Clarias Gariepinus ◽

Fish Farming ◽

Progeny Test ◽

African Catfish ◽

Mendelian Segregation ◽

Qpcr Method ◽

Pcr Method ◽

Real Time Qpcr ◽

Transgene Transmission

Produktivitas ikan budidaya dapat ditingkatkan melalui teknologi transgenesis. Populasi ikan lele transgenik cepat tumbuh telah dihasilkan dan karakter biologisnya telah diketahui. Namun informasi zigositas ikan lele transgenik perlu ditelaah lebih lanjut. Penelitian ini bertujuan untuk mengidentifikasi zigositas ikan lele transgenik F-2. Zigositas ikan lele transgenik diidentifikasi dengan menggunakan metode real-time qPCR (RT-qPCR) dan uji progeni. Identifikasi zigositas melalui uji progeni, dilakukan dengan mendeteksi transgen (PhGH) pada individu-individu F-3 hasil persilangan transgenik F-2 dengan non-transgenik. Hasil penelitian menunjukkan bahwa zigositas pada ikan lele transgenik F-2 dapat diidentifikasi dengan menggunakan metode RT-qPCR. Semua ikan transgenik F-2 adalah heterozigot, dengan nilai 2-Ct yang hampir sama tiap individu F-2, yaitu berkisar 0,80-0,99. Identifikasi zigositas dengan metode RT-qPCR menunjukkan hasil yang sama dengan uji progeni, semua transgenik F-2 tidak menghasilkan 100% anakan F-3 positif transgen. Pada uji progeni, transmisi transgen pada penelitian ini tidak mengikuti hukum segregasi Mendel, dengan kisaran sebesar 5%-40%.Fish farming productivity can be increased by transgenesis technology. On the previous study, transgenic African catfish population fast growing has been produced and its biological characters has been known. However information of transgenic zygosity of catfish should be examined. The aim of this study was to identify the zygosity of F-2 transgenic African catfish. The zygosity of F-2 transgenic was identified by real time-qPCR (RT-qPCR) method and progeny test. Further, identification of zygosity F-2 transgenic African catfish was confirmed by progeny test, while F-2 transgenic African catfish was mated with non-transgenic. Identification of zygosity F-2 transgenic was conducted by detection PhGH gene (transgene) in F-3 transgenic African catfish population. Transgene transmission was evaluated by PCR method. The result showed that the zygosity F-2 transgenic African catfish could be identified by RT-qPCR method. All F-2 transgenic African catfish were heterozygous, where as the 2-Ct value was almost same for all individual, which ranges from 0.80 to 0.99. The result of zygosity identification using RT-qPCR method was as same as that of progeny test. In the progeny test, transgene transmission in this study was non-Mendelian segregation, with ranges of 5%-40%.

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Author Correction: A simple and robust real-time qPCR method for the detection of PIK3CA mutations

Scientific Reports ◽

10.1038/s41598-021-95597-0 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Virginia Alvarez-Garcia ◽

Clare Bartos ◽

Ieva Keraite ◽

Urmi Trivedi ◽

Paul M. Brennan ◽

...

Keyword(s):

Real Time ◽

Pik3ca Mutations ◽

Qpcr Method ◽

Real Time Qpcr

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Establishment of a Duplex Real-time qPCR Method for Detection of Salmonella Spp. And Serratia Fonticola in Fishmealestablishment of a Duplex Real-time qPCR Method for Detection of Salmonella Spp. And Serratia Fonticola in Fishmeal

10.21203/rs.3.rs-78082/v1 ◽

2020 ◽

Author(s):

Jinghua Ruan ◽

Wujun Wang ◽

Tiying Zhang ◽

Teng Zheng ◽

Jing Zheng ◽

...

Keyword(s):

Real Time ◽

Limit Of Detection ◽

Bacterial Species ◽

High Specificity ◽

Clinical Samples ◽

Amplification Efficiency ◽

Salmonella Spp ◽

Specificity And Sensitivity ◽

Qpcr Method ◽

Real Time Qpcr

Abstract Salmonella spp. is a high-risk bacterial pathogen that is monitored in imported animal-derived feedstuffs. Serratiafonticola is the bacterial species most frequently confused with Salmonella spp. in traditional identification methods based on biochemical characteristics, which are time-consuming and labor-intensive, and thus unsuitable for daily inspection and quarantine work. In this study, we established a duplex real-time qPCR method with invA- and gyrB-specific primers and probes corresponding to Salmonella spp. and S. fonticola. The method could simultaneously detect both pathogens in imported feedstuffs, with a minimum limit of detection for Salmonella spp. and S. fonticola of 197 copies/SL and 145 copies/SL, respectively (correlation coefficient R2 = 0.999 in both cases). The amplification efficiency for Salmonella spp. and S. fonticola was 98.346% and 96.49%, respectively. Detection of clinical samples was consistent with method GB/T 13091-2018, and all seven artificially contaminated imported feed samples were positively identified. Thus, the developed duplex real-time qPCR assay displays high specificity and sensitivity, and can be used for the rapid and accurate detection of genomic DNA from Salmonella spp. and S. fonticola within hours. This represents a significant improvement in the efficiency of detection of both pathogens in imported feedstuffs.

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Establishment of a duplex real-time qPCR method for detection of Salmonella spp. and Serratia fonticola in fishmeal

AMB Express ◽

10.1186/s13568-020-01144-x ◽

2020 ◽

Vol 10 (1) ◽

Author(s):

Jinghua Ruan ◽

Wujun Wang ◽

Tiyin Zhang ◽

Teng Zheng ◽

Jing Zheng ◽

...

Keyword(s):

Real Time ◽

Limit Of Detection ◽

Bacterial Species ◽

Bacterial Pathogen ◽

High Specificity ◽

Amplification Efficiency ◽

Salmonella Spp ◽

Specificity And Sensitivity ◽

Qpcr Method ◽

Real Time Qpcr

AbstractSalmonella spp. is a high-risk bacterial pathogen that is monitored in imported animal-derived feedstuffs. Serratia fonticola is the bacterial species most frequently confused with Salmonella spp. in traditional identification methods based on biochemical characteristics, which are time-consuming and labor-intensive, and thus unsuitable for daily inspection and quarantine work. In this study, we established a duplex real-time qPCR method with invA- and gyrB-specific primers and probes corresponding to Salmonella spp. and S. fonticola. The method could simultaneously detect both pathogens in imported feedstuffs, with a minimum limit of detection for Salmonella spp. and S. fonticola of 197 copies/μL and 145 copies/μL, respectively (correlation coefficient R2 = 0.999 in both cases). The amplification efficiency for Salmonella spp. and S. fonticola was 98.346% and 96.49%, respectively. Detection of fishmeal was consistent with method GB/T 13091-2018, and all seven artificially contaminated imported feed samples were positively identified. Thus, the developed duplex real-time qPCR assay displays high specificity and sensitivity, and can be used for the rapid and accurate detection of genomic DNA from Salmonella spp. and S. fonticola within hours. This represents a significant improvement in the efficiency of detection of both pathogens in imported feedstuffs.

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Quantification of Airborne Influenza and Avian Influenza Virus in a Wet Poultry Market using a Filter/Real-time qPCR Method

Aerosol Science and Technology ◽

10.1080/02786820802621232 ◽

2009 ◽

Vol 43 (4) ◽

pp. 290-297 ◽

Cited By ~ 24

Author(s):

Pei-Shih Chen ◽

Chien Kun Lin ◽

Feng Ta Tsai ◽

Chun-Yuh Yang ◽

Chien-Hung Lee ◽

...

Keyword(s):

Influenza Virus ◽

Avian Influenza ◽

Avian Influenza Virus ◽

Real Time ◽

Qpcr Method ◽

Real Time Qpcr

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Bioconversion of Dried Leaves from Algerian Date Palm (Phoenix dactylifera L.) to Biogas by Anaerobic Digestion

International Journal of Engineering Research in Africa ◽

10.4028/www.scientific.net/jera.41.131 ◽

2019 ◽

Vol 41 ◽

pp. 131-144

Author(s):

Mohammed Djaafri ◽

Slimane Kalloum ◽

Ahmed Elamine Soulimani ◽

Mostefa Khelafi

Keyword(s):

Anaerobic Digestion ◽

Date Palm ◽

Biogas Production ◽

Stable Process ◽

Production Capacity ◽

Oxygen Demand ◽

Phoenix Dactylifera ◽

Total Alkalinity ◽

Process Stability ◽

Biogas Yield

This work aimed to study anaerobic digestion of a new substrate made from the waste of Algerian date palm (Phoenix dactyliferaL.) in order to evaluate its biogas production capacity. The lignocellulosic component biodegradability and the process stability were also tested. Two reactors were launched with the same concentration of 20g total solid (TS)/L. A physical pre-treatment (grinding) of dried leaves lowered the substrate size below 2 mm. The theoretical methane yield (499 - 538 mL CH4/g of Volatile Solids (VS)) indicated a high substrate methanogen potential. The tested methane and biogas yield reached 130±6 mL CH4/gVS and 197±6 mL/g VS respectively. The biodegradability through the volatiles solid reduction was 29%. The total alkalinity (TA) value increased from the beginning to the end, due to the substrate nature which gave a high buffering capacity of the medium. During the stable process, a higher Chemical Oxygen Demand (COD) removal was reached with a value of 77.58%. The majority of the biogas yield was generated during the same period, which confirmed that the anaerobic digestion process stability is indispensable. Finally, this study was an opportunity to better exploit local by-products for the bioenergy production.

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A simple and robust real-time qPCR method for the detection of PIK3CA mutations

Scientific Reports ◽

10.1038/s41598-018-22473-9 ◽

2018 ◽

Vol 8 (1) ◽

Cited By ~ 8

Author(s):

Virginia Alvarez-Garcia ◽

Clare Bartos ◽

Ieva Keraite ◽

Urmi Trivedi ◽

Paul M. Brennan ◽

...

Keyword(s):

Real Time ◽

Pik3ca Mutations ◽

Qpcr Method ◽

Real Time Qpcr

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Establishment and Application of a duplex real-time qPCR method for detection ofSalmonellaspp. andSerratia fonticolain imported feedstuffs

10.1101/324798 ◽

2018 ◽

Author(s):

Jing hua Ruan ◽

Wu jun Wang ◽

Quan yang Bai ◽

Ti yin Zhang ◽

Teng zheng ◽

...

Keyword(s):

Real Time ◽

Limit Of Detection ◽

Bacterial Species ◽

High Specificity ◽

Accurate Method ◽

Clinical Samples ◽

Amplification Efficiency ◽

Specificity And Sensitivity ◽

Qpcr Method ◽

Real Time Qpcr

AbstractSalmonellaspp. is a high-risk bacterial pathogen that is monitored in imported animal-derived feedstuffs.Serratia fonticolais the bacterial species most frequently confused withSalmonellaspp. in traditional identification methods based on biochemical characteristics, which are time-consuming and labor-intensive, and thus unsuitable for daily inspection and quarantine work. In this study, we established a duplex real-time qPCR method withinvA-andgyrB-specific primers and probes corresponding toSalmonellaspp. andS. fonticola. The method could simultaneously detect both pathogens in imported feedstuffs, with a minimum limit of detection forSalmonellaspp. andS. fonticolaof 197 copies/μL and 145 copies/μL, respectively (correlation coefficient R2= 0.999 in both cases). The amplification efficiency forSalmonellaspp. andS. fonticolawas 98.346% and 96.49%, respectively. Detection of clinical samples was consistent with method GB/T 13091-2002, and all 20 artificially contaminated imported feed samples were positively identified. Thus, the developed duplex real-time qPCR assay displays high specificity and sensitivity, and can be used for the rapid and accurate detection of genomic DNA fromSalmonellaspp. andS. fonticolawithin hours. This represents a significant improvement in the efficiency of detection of both pathogens in imported feedstuffs.ImportanceImported feedstuffs must be tested for pathogenicSalmonellaspecies that represent a biological hazard. Variousnon-Salmonellacolony-forming species belong toEnterobacteriaceae,andSerratia fonticolaforms colonies of similar color and morphology toSalmonellaspp., leading to confusion in daily quarantine tests. Traditional methods based on biochemical and serological characteristics are cumbersome and labor-intensive, and unable to fully support current quarantine testing demands. Thus, there is an urgent need to develop a rapid and accurate method for the effective identification of these pathogens. The duplex real-time qPCR method established herein can rapidly identifySalmonellaspp. andS. fonticola, and has great potential for application to feed safety and prevention of exterior pathogens.

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