title: Development and evaluation of loop-mediated isothermal amplification assay for rapid and inexpensive detection of cytomegalovirus DNA in vitreous specimens from suspected cases of viral retinitis.
abstract: A loop-mediated isothermal amplification (LAMP) assay for the rapid detection of cytomegalovirus (CMV) was developed and evaluated. The LAMP assay specifically amplified only CMV DNA, and no cross-reactivity with the DNA of herpes simplex virus type 1, varicella-zoster virus, adenovirus, Aspergillus flavus, or Staphylococcus aureus was observed. The sequences of the LAMP assay-positive CMV products were perfectly (100%) matched with the CMV sequence deposited in the GenBank database. The sensitivity of the LAMP assay was found to be 10 copies/microl of CMV DNA. Vitreous samples from 40 patients with suspected retinitis were subjected to LAMP and real-time PCR for the detection of CMV. Of 40 patients with suspected viral retinitis, 10 tested positive for CMV by the real-time PCR and LAMP assays. A 100% concordance was observed between the results of the two methods. The LAMP assay is a rapid, highly specific, and sensitive method for the diagnosis of retinitis caused by CMV.
authors.affiliation: Jhaveri Microbiology Centre, L. V. Prasad Eye Institute, Banjara Hills, Road No. 2, Hyderabad 500 034, India. ashokkumar@lvpei.org
authors.forename: Ashok Kumar
authors.initials: AK
authors.lastname: Reddy
authors.forename: Praveen Kumar
authors.initials: PK
authors.lastname: Balne
authors.forename: Rajeev Kumar
authors.initials: RK
authors.lastname: Reddy
authors.forename: Annie
authors.initials: A
authors.lastname: Mathai
authors.forename: Inderjeet
authors.initials: I
authors.lastname: Kaur
mesh_terms: D003587:Cytomegalovirus / Q000235:genetics / Q000302:isolation & purification*; D017726:Cytomegalovirus Retinitis / Q000175:diagnosis* / Q000821:virology; D004279:DNA, Viral / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D012680:Sensitivity and Specificity; D013696:Temperature; D014773:Virology / Q000379:methods*; D014822:Vitreous Body / Q000821:virology*
chemical_list: D004279:DNA, Viral