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Rapid detection and subtyping of human papillomaviruses in condyloma acuminatum using loop-mediated isothermal amplification with hydroxynaphthol blue dye.
Br J Biomed Sci. 2018 Jul; 75(3):110-115.BJ

Abstract

Objective Condyloma acuminatum (CA) is a common, viral, sexually transmitted disease worldwide. Human papillomavirus (HPV) genotyping has important clinical implications for the treatment of CA. We developed a loop-mediated isothermal amplification (LAMP) method for the detection of HPV. Methods We collected 294 cervical scrape samples, including 30 HPV-6-positive, 30 HPV-11-positive, 22 HPV-16-positive, 20 HPV-42-positve, 30 HPV-43-positive, 20 HPV-44-positive and 142 HPV-negative samples. Tissues from 40 patients with a pathological diagnosis of CA were paraffin-embedded and analyzed by LAMP and Luminex. Hydroxynaphthol blue (HNB) and electrophoresis were used to detect the results of LAMP. Results LAMP and Luminex systems were compared in detecting six subtypes of HPV. LAMP reactions were specific for each subtype. The sensitivity of LAMP for HPV-6, as determined by the HNB indicator assay, was 1000 copies/tube. The kappa value between the two methods was 0.98 (HPV-6), 0.94 (HPV-11), 0.89 (HPV-43), 0.87 (HPV-42) 0.79 (HPV-16) and 0.68 (HPV-44). Among the 142 HPV-negative samples determined by the Luminex assay, HPV-6 was detected in eight and HPV-11 in one by LAMP. Among the 40 CA samples, the results of LAMP and Luminex were in agreement in 38 (95%). Conclusion The results of this study indicated that the LAMP assay with HNB is superior to the Luminex method in terms of sensitivity and specificity. The specificity of LAMP was 100% and the sensitivity of LAMP was 1000 copies/tube using HNB. LAMP is therefore a useful, quick and accurate method for the clinical diagnosis of HPV subtypes.

Authors+Show Affiliations

a Department of Central Laboratory , Shanghai Tenth People's Hospital, Tongji University , Shanghai , China. b Department of Laboratory Medicine , Shanghai Tenth People's Hospital, Tongji University , Shanghai , China. c Department of Laboratory Medicine , Shanghai Skin Disease Hospital, Tongji University , Shanghai , China.a Department of Central Laboratory , Shanghai Tenth People's Hospital, Tongji University , Shanghai , China. b Department of Laboratory Medicine , Shanghai Tenth People's Hospital, Tongji University , Shanghai , China.a Department of Central Laboratory , Shanghai Tenth People's Hospital, Tongji University , Shanghai , China. b Department of Laboratory Medicine , Shanghai Tenth People's Hospital, Tongji University , Shanghai , China.d Department of Laboratory Medicine , Huashan Hospital, Fudan University , Shanghai , China.e Department of Radiation Oncology , Shanghai Tenth People's Hospital, Tongji University , Shanghai , China.a Department of Central Laboratory , Shanghai Tenth People's Hospital, Tongji University , Shanghai , China. b Department of Laboratory Medicine , Shanghai Tenth People's Hospital, Tongji University , Shanghai , China. c Department of Laboratory Medicine , Shanghai Skin Disease Hospital, Tongji University , Shanghai , China.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

29537347

Citation

Zhong, Q, et al. "Rapid Detection and Subtyping of Human Papillomaviruses in Condyloma Acuminatum Using Loop-mediated Isothermal Amplification With Hydroxynaphthol Blue Dye." British Journal of Biomedical Science, vol. 75, no. 3, 2018, pp. 110-115.
Zhong Q, Li K, Chen D, et al. Rapid detection and subtyping of human papillomaviruses in condyloma acuminatum using loop-mediated isothermal amplification with hydroxynaphthol blue dye. Br J Biomed Sci. 2018;75(3):110-115.
Zhong, Q., Li, K., Chen, D., Wang, H., Lin, Q., & Liu, W. (2018). Rapid detection and subtyping of human papillomaviruses in condyloma acuminatum using loop-mediated isothermal amplification with hydroxynaphthol blue dye. British Journal of Biomedical Science, 75(3), 110-115. https://doi.org/10.1080/09674845.2017.1411864
Zhong Q, et al. Rapid Detection and Subtyping of Human Papillomaviruses in Condyloma Acuminatum Using Loop-mediated Isothermal Amplification With Hydroxynaphthol Blue Dye. Br J Biomed Sci. 2018;75(3):110-115. PubMed PMID: 29537347.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Rapid detection and subtyping of human papillomaviruses in condyloma acuminatum using loop-mediated isothermal amplification with hydroxynaphthol blue dye. AU - Zhong,Q, AU - Li,K, AU - Chen,D, AU - Wang,H, AU - Lin,Q, AU - Liu,W, Y1 - 2018/03/14/ PY - 2018/3/15/pubmed PY - 2019/8/1/medline PY - 2018/3/15/entrez KW - HPV KW - LAMP KW - Loop-mediated isothermal amplification KW - human papillomavirus SP - 110 EP - 115 JF - British journal of biomedical science JO - Br. J. Biomed. Sci. VL - 75 IS - 3 N2 - Objective Condyloma acuminatum (CA) is a common, viral, sexually transmitted disease worldwide. Human papillomavirus (HPV) genotyping has important clinical implications for the treatment of CA. We developed a loop-mediated isothermal amplification (LAMP) method for the detection of HPV. Methods We collected 294 cervical scrape samples, including 30 HPV-6-positive, 30 HPV-11-positive, 22 HPV-16-positive, 20 HPV-42-positve, 30 HPV-43-positive, 20 HPV-44-positive and 142 HPV-negative samples. Tissues from 40 patients with a pathological diagnosis of CA were paraffin-embedded and analyzed by LAMP and Luminex. Hydroxynaphthol blue (HNB) and electrophoresis were used to detect the results of LAMP. Results LAMP and Luminex systems were compared in detecting six subtypes of HPV. LAMP reactions were specific for each subtype. The sensitivity of LAMP for HPV-6, as determined by the HNB indicator assay, was 1000 copies/tube. The kappa value between the two methods was 0.98 (HPV-6), 0.94 (HPV-11), 0.89 (HPV-43), 0.87 (HPV-42) 0.79 (HPV-16) and 0.68 (HPV-44). Among the 142 HPV-negative samples determined by the Luminex assay, HPV-6 was detected in eight and HPV-11 in one by LAMP. Among the 40 CA samples, the results of LAMP and Luminex were in agreement in 38 (95%). Conclusion The results of this study indicated that the LAMP assay with HNB is superior to the Luminex method in terms of sensitivity and specificity. The specificity of LAMP was 100% and the sensitivity of LAMP was 1000 copies/tube using HNB. LAMP is therefore a useful, quick and accurate method for the clinical diagnosis of HPV subtypes. SN - 0967-4845 UR - https://www.unboundmedicine.com/medline/citation/29537347/Rapid_detection_and_subtyping_of_human_papillomaviruses_in_condyloma_acuminatum_using_loop_mediated_isothermal_amplification_with_hydroxynaphthol_blue_dye_ L2 - https://medlineplus.gov/genitalwarts.html DB - PRIME DP - Unbound Medicine ER -