Development of loop-mediated isothermal amplification method for detecting Wuchereria bancrofti DNA in human blood and vector mosquitoes

• Published in: Parasitology international Vol. 60; no. 4; pp. 493 - 497
• Main Authors: Takagi, Hidekazu, Itoh, Makoto, Kasai, Shinji, Yahathugoda, Thishan C, Weerasooriya, Mirani V, Kimura, Eisaku
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier Ireland Ltd 01-12-2011
• Subjects: Animals; Base Sequence; blood; Brugia - genetics; Brugia malayi; Culex - genetics; Culex quinquefasciatus; Dirofilaria - genetics; Dirofilaria immitis; DNA; DNA, Helminth - analysis; DNA, Helminth - genetics; Filariasis - blood; Filariasis - diagnosis; Filariasis - parasitology; Filarioidea; Gastroenterology and Hepatology; human diseases; Humans; Infectious Disease; loop-mediated isothermal amplification; Loop-mediated isothermal amplification (LAMP); Molecular Sequence Data; monitoring; Nucleic Acid Amplification Techniques; parasitology; patients; polymerase chain reaction; Repetitive Sequences, Nucleic Acid - genetics; Sensitivity and Specificity; Wuchereria bancrofti; Wuchereria bancrofti - genetics; Wuchereria bancrofti - isolation & purification; Loop-mediated isothermal amplification (LAMP); Wuchereria bancrofti; Culex quinquefasciatus
• ISSN: 1383-5769; 1873-0329
• DOI: 10.1016/j.parint.2011.08.018

Abstract We have developed loop-mediated isothermal amplification (LAMP) method to detect Wuchereria bancrofti DNA. The sensitivity and specificity of LAMP method were equivalent to those of PCR method which detects Ssp I repeat sequence in W. bancrofti genomic DNA: both methods detected one thousandth of W. bancrofti DNA from one microfilaria (Mf), and did not cross-react with DNAs of Brugia malayi , B. pahangi , Dirofilaria immitis , human and Culex quinquefasciatus . We also examined the sensitivity of LAMP using the mimic samples of patient's blood or blood-fed mosquitoes containing one W. bancrofti Mf per sample. The LAMP method was able to detect W. bancrofti DNA in 1000 μl of blood or in a pool of 60 mosquitoes, indicating its usefulness in detecting/monitoring W. bancrofti infection in humans and vector mosquitoes in endemic areas.