title: Coupling electrochemical response of a DNA biosensor with PCR for Neisseria gonorrhoeae detection.
abstract: Early diagnosis of gonococcal infections is important with regard to a patient's health and stage of infection. In this context, we report the development of an opa-gene-based electrochemical DNA biosensor for detection of Neisseria gonorrhoeae by monitoring redox peak of methylene blue indicator. The fabricated biosensor has been shown to be highly sensitive and specific when evaluated with complementary, non-complementary, and 1-base mismatch DNA sequences and polymerase chain reaction (PCR) amplified products (amplicons) of standard strain of N. gonorrhoeae (ATCC49226). The biosensor has been further evaluated using amplicons of known positive and negative clinical samples, and cut-off for positives has been determined using receiver operating characteristic curve. The sensitivity (SN), specificity (SP), positive predictive value, and negative predictive value of the biosensor have been found to be 96.2%, 88.2%, 92.6%, and 93.8%, respectively. We conclude that the combination of PCR amplification with electrochemical detection shows distinct advantage of high SN and increased SP for gonococcal detection.
authors.affiliation: Department of Microbiology, All India Institute of Medical Sciences, New Delhi, India.
authors.forename: Rachna
authors.initials: R
authors.lastname: Verma
authors.affiliation: Department of Microbiology, All India Institute of Medical Sciences, New Delhi, India. Electronic address: seemalsood@rediffmail.com.
authors.forename: Seema
authors.initials: S
authors.lastname: Sood
authors.affiliation: DST Centre on Biomolecular electronics, Biomedical Instrumentation Section, National Physical Laboratory (Council of Scientific & Industrial Research), Dr. K.S. Krishnan Marg, New Delhi-110012, India; School of Biotechnology, Guru Gobind Singh Indraprastha University, Sector 16 C, Dwarka, Delhi, 110075 India.
authors.forename: Renu
authors.initials: R
authors.lastname: Singh
authors.affiliation: DST Centre on Biomolecular electronics, Biomedical Instrumentation Section, National Physical Laboratory (Council of Scientific & Industrial Research), Dr. K.S. Krishnan Marg, New Delhi-110012, India.
authors.forename: Gajjala
authors.initials: G
authors.lastname: Sumana
authors.affiliation: Regional STD Teaching, Training and Research Centre, VMMC & SJH, New Delhi, India.
authors.forename: Manju
authors.initials: M
authors.lastname: Bala
authors.affiliation: Department of Dermatology and Venereology, All India Institute of Medical Sciences, New Delhi, India.
authors.forename: Vinod K
authors.initials: VK
authors.lastname: Sharma
authors.affiliation: Department of Microbiology, All India Institute of Medical Sciences, New Delhi, India.
authors.forename: Jyotish C
authors.initials: JC
authors.lastname: Samantaray
authors.affiliation: Department of Biostatistics, All India Institute of Medical Sciences, New Delhi, India.
authors.forename: Ravindra M
authors.initials: RM
authors.lastname: Pandey
authors.affiliation: DST Centre on Biomolecular electronics, Biomedical Instrumentation Section, National Physical Laboratory (Council of Scientific & Industrial Research), Dr. K.S. Krishnan Marg, New Delhi-110012, India; Department of Biotechnology, Delhi Technological University, New Delhi-110042, India.
authors.forename: Bansi D
authors.initials: BD
authors.lastname: Malhotra
mesh_terms: D000293:Adolescent; D000328:Adult; D015374:Biosensing Techniques / Q000379:methods*; D003955:Diagnostic Tests, Routine / Q000379:methods*; D042241:Early Diagnosis; D055664:Electrochemical Techniques / Q000379:methods*; D005260:Female; D006069:Gonorrhea / Q000175:diagnosis*; D006801:Humans; D008297:Male; D008875:Middle Aged; D009344:Neisseria gonorrhoeae / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods*; D011237:Predictive Value of Tests; D012680:Sensitivity and Specificity; D055815:Young Adult
keywords: Diagnosis; Electrochemical DNA biosensor; Gonorrhoea; Methylene Blue; Neisseria gonorrhoeae; PCR