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.
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.
Reddy
Ashok Kumar
AK
Jhaveri Microbiology Centre, L. V. Prasad Eye Institute, Banjara Hills, Road No. 2, Hyderabad 500 034, India. ashokkumar@lvpei.org
Balne
Praveen Kumar
PK
Reddy
Rajeev Kumar
RK
Mathai
Annie
A
Kaur
Inderjeet
I
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*
D004279:DNA, Viral