Rapid detection of human immunodeficiency virus type 1 group M by a reverse transcription-loop-mediated isothermal amplification assay.
A rapid one-step reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay targeting the pol-integrase gene was developed to detect human immunodeficiency virus type 1 (HIV-1) group M. This HIV-1 RT-LAMP assay is simple and rapid, and amplification can be completed within 35min under isothermal conditions at 60 degrees C. The 100% detection limit of HIV-1 RT-LAMP was determined using a standard strain (WHO HIV-1 [97/656]) in octuplicate and found to be 120 copies/ml. The RT-LAMP assay was evaluated for use for clinical diagnosis using plasma samples collected from 57 HIV-1-infected and 40 uninfected individuals in Cameroon, where highly divergent HIV-1 strains are prevalent. Of the 57 samples from infected individuals, 56 harbored group-M HIV-1 strains, such as subtypes A, B, G, F2, and circulating recombinant forms (CRFs) _01, _02, _09, _11, _13; all were RT-LAMP positive. One sample harboring group-O HIV-1 and the 40 HIV-1-uninfected samples were RT-LAMP negative. These findings indicate that HIV-1 RT-LAMP can detect HIV-1 group-M RNA from plasma samples rapidly and with high sensitivity and specificity. These data also suggest that this RT-LAMP assay can be useful for confirming HIV diagnosis, particularly in resource-limited settings.
Hosaka
Norimitsu
N
Department of Viral Infection and International Health, Kanazawa University, Takaramachi, Japan.
Ndembi
Nicaise
N
Ishizaki
Azumi
A
Kageyama
Seiji
S
Numazaki
Kei
K
Ichimura
Hiroshi
H
D002163:Cameroon; D005838:Genotype; D015658:HIV Infections / Q000175:diagnosis* / Q000821:virology*; D019427:HIV Integrase / Q000235:genetics; D015497:HIV-1 / Q000145:classification* / Q000302:isolation & purification*; D006801:Humans; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D012680:Sensitivity and Specificity; D054302:pol Gene Products, Human Immunodeficiency Virus / Q000235:genetics
D054302:pol Gene Products, Human Immunodeficiency Virus; D019427:HIV Integrase