scholarly journals Standardization of Loop mediated isothermal amplification for detection of D. nodosus and F. necrophorum causing footrot in sheep and goats

2021 ◽  
Author(s):  
kavitha madineni ◽  
N. Vinod Kumar
Keyword(s):  
Detection Limit ◽  
16S Rrna ◽  
16S Rrna Gene ◽  
Rapid Detection ◽  
Statistical Significance ◽  
Isothermal Amplification ◽  
Rrna Gene ◽  
Sheep And Goats ◽  
Loop Mediated Isothermal Amplification ◽  
High Statistical Significance

Abstract The Loop Mediated Isothermal Amplification (LAMP) was standardized for rapid detection of D. nodosus and F. necrophorum. A total of 250 foot swabs (200) were screened from sheep and (50) were from goats from different districts of Rayalaseema viz., Chittoor, Nellore, Kadapa, Anantapur. Out of 250 samples 75 (30.0%) and 85 (34.0%) were positive for D. nodosus and F. necrophorum respectively. All the 250 samples were screened individually for both the organisms by LAMP. Among them, 104 (41.6%) were found to be positive for D. nodosus and 120 (48.0%) were positive for F. necrophorum. The efficacy of LAMP in terms of sample DNA detection limit was compared with the PCR by using standard dilutions of DNA extracted from D. nodosus and F. necrophorum cultures. The detection limit was found to be higher than PCR for both the organisms. The sensitivity of LAMP is compared with PCR by targeting 16S rRNA gene of D. nodosus and lktA gene of F. necrophorum. In case of D. nodosus, out of 250 samples, 75 (30.0%) were positive by PCR and 104 (41.6%) were positive by LAMP. Among 250 samples, 85 (34.0%) were positive by PCR and 120 (48.0%) were positive by LAMP in case of F. necrophorum. The LAMP was found to be more sensitive than PCR in detecting the organisms with high statistical significance.

Rapid detection of ammonia-oxidizing bacteria in activated sludge based on 16S-rRNA gene by using PCR and fluorometry

2002 ◽  
Vol 7 (5) ◽  
pp. 323-326
Author(s):  
Motohiko Hikuma ◽  
Masanori Nakajima ◽  
Toshiaki Hirai ◽  
Hiroshi Matsuoka
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Activated Sludge ◽  
Rapid Detection ◽  
Rrna Gene ◽  
Ammonia Oxidizing Bacteria

scholarly journals Rapid Detection of Subtype H10N8 Influenza Virus by One-Step Reverse Transcription–Loop-Mediated Isothermal Amplification Methods

2015 ◽  
Vol 53 (12) ◽  
pp. 3884-3887 ◽  
Author(s):  
Hongmei Bao ◽  
Xiaoxiao Feng ◽  
Yong Ma ◽  
Jianzhong Shi ◽  
Yuhui Zhao ◽  
...  
Keyword(s):  
Influenza Virus ◽  
Detection Limit ◽  
Reverse Transcription ◽  
Rapid Detection ◽  
High Sensitivity ◽  
Isothermal Amplification ◽  
Loop Mediated Isothermal Amplification ◽  
Live Poultry ◽  
One Step ◽  
Live Poultry Markets

We developed hemagglutinin- and neuraminidase-specific one-step reverse transcription–loop-mediated isothermal amplification assays for detecting the H10N8 virus. The detection limit of the assays was 10 copies of H10N8 virus, and the assays did not amplify nonspecific RNA. The assays can detect H10N8 virus from chicken samples with high sensitivity and specificity, and they can serve as an effective tool for detecting and monitoring H10N8 virus in live poultry markets.

scholarly journals PCR-Based Identification of Hyperthermophilic Archaea of the Family Thermococcaceae

2004 ◽  
Vol 70 (9) ◽  
pp. 5701-5703 ◽  
Author(s):  
Galina B. Slobodkina ◽  
Nikolai A. Chernyh ◽  
Alexander I. Slobodkin ◽  
Irina V. Subbotina ◽  
Elizaveta A. Bonch-Osmolovskaya ◽  
...  
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Rapid Detection ◽  
Pcr Amplification ◽  
Hyperthermophilic Archaea ◽  
Rrna Gene ◽  
Detection And Identification ◽  
The Family

ABSTRACT A method for rapid detection and identification of hyperthermophilic archaea of the family Thermococcaceae based on PCR amplification of 16S rRNA gene fragments with primers TcPc 173F (5′-TCCCCCATAGGYCTGRGGTACTGGAAGGTC-3′) and TcPc 589R (5′-GCCGTGRGATTTCGCCAGGGACTTACGGGC-3′) was developed and used for identification of new isolates.

Differentiation Of Clarias Batrachus, C. Gariepinus And Heteropneustes Fossilis By Pcr-Sequencing Of Mitochondrial 16s Rrna Gene

2015 ◽  
Vol 41 (1) ◽  
pp. 51-58
Author(s):  
Mohammad Shamimul Alam ◽  
Hawa Jahan ◽  
Rowshan Ara Begum ◽  
Reza M Shahjahan
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Fish Larva ◽  
16S Rrna Genes ◽  
Rrna Genes ◽  
Rrna Gene ◽  
Valuable Insight ◽  
Multiple Sequence ◽  
Pcr Rflp ◽  
Alternative Means

Heteropneustesfossilis, Clariasbatrachus and C. gariepinus are three major catfishes ofecological and economic importance. Identification of these fish species becomes aproblem when the usual external morphological features of the fish are lost or removed,such as in canned fish. Also, newly hatched fish larva is often difficult to identify. PCRsequencingprovides accurate alternative means of identification of individuals at specieslevel. So, 16S rRNA genes of three locally collected catfishes were sequenced after PCRamplification and compared with the same gene sequences available from othergeographical regions. Multiple sequence alignment of the 16S rRNA gene fragments ofthe catfish species has revealed polymorphic sites which can be used to differentiate thesethree species from one another and will provide valuable insight in choosing appropriaterestriction enzymes for PCR-RFLP based identification in future. Asiat. Soc. Bangladesh, Sci. 41(1): 51-58, June 2015

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