Development and evaluation of loop-mediated isothermal amplifiation for rapid detection of Nosema ceranae in honeybee

Journal Title: Asian Pacific Journal of Tropical Disease - Year 2016, Vol 6, Issue 12

Abstract

Objective: To develop loop-mediated isothermal amplification (LAMP) to detect Nosema ceranae (N. ceranae) in honeybee samples. Methods: LAMP primers were designed recognizing six distinct fragments of 16s rRNA gene and LAMP reaction was determined by optimizing the concentration of reagents, such as forward inner primer and backward inner primer, deoxynucleoside triphosphate and betaine, time and temperature. Ten-fold serial dilutions of DNA were used to determine the detection limit and accuracy using both LAMP and PCR tests. Results: LAMP required 1.2 µmol/L of forward inner primer and backward inner primer primers, 0.2 µmol/L of forward outer primers and backward outer primer, 2 µmol/L of Mg2+, 0.6 mol/L of betaine, 0.6 µmol/L of deoxynucleoside triphosphate, 4.8 IU of Bst DNA polymerase and 30 ng of DNA. The optimal temperature was 63 °C and after a 40-min incubation time, a clearly ladder-like pattern of LAMP product appeared in the gel electrophoresis. LAMP appeared more sensitive than a standard PCR in detection of N. ceranae. Conclusions: LAMP gave a good results and it could be an alternative diagnostic tool instead of PCR to detect N. ceranae infection in honeybee.

Authors and Affiliations

Keywords

Related Articles

Immunomodulatory activity of an aqueous extract of Polygonum minus Huds on Swiss albino mice using carbon clearance assay

Objective: To explore the immunomodulatory properties of aqueous extract of Polygonum minus Huds (P. minus), on swiss albino mice using carbon clearance assay. Methods: The study was performed using in-vivo carbon cleara...

Some discussions on imported dengue virus and use of adjunctive recombinant activated factor VIIa in dengue

Dengue virus infection is common in tropical world and becomes the big public health threat in tropical medicine. The changing of its epidemiology is very interesting. The good transportation is an important consideratio...

Malaria in the Andaman and Nicobar Islands: Challenges and opportunities for elimination

Malaria has been endemic to Andaman and Nicobar Archipelago for over a century. However, the cases were observed to decline during the past few years. Plasmodium vivax and Plasmodium falciparum are the predominant malari...

Bacteria etiological agents causing respiratory tract infections in children and their resistance patterns to a panel of ten antibiotics

Objective: To study the bacteria etiological agents of respiratory tract infection among 280 school children in South East Nigeria, and to determine their antimicrobial resistance patterns to a panel of ten antibiotics....

Download PDF file
  • EP ID EP226993
  • DOI 10.1016/S2222-1808(16)61163-5
  • Views 69
  • Downloads 0

How To Cite

(2016). Development and evaluation of loop-mediated isothermal amplifiation for rapid detection of Nosema ceranae in honeybee. Asian Pacific Journal of Tropical Disease, 6(12), 952-956. https://europub.co.uk/articles/-A-226993