O, evaluation of the efficacy of laboratory made lamp method indetection of human papilloma virus

Human Papillomaviruses (HPV) are DNA viruses that cause skin and mucous membrane infections in humans. HPV infection ranks first among the most common sexually transmitted infections. Today, more than 200 HPV types have been identified and persistent infection caused by oncogenic HPV types known as high risk (HR) types can cause precancerous lesions and cancer. Among these, HPV 16 and HPV 18 types have the highest carcinogenic potential. Cervical cancer ranks first among cancers caused by HR HPV types and the majority of cervical cancer cases are caused by HPV 16 and HPV 18 infections. Appropriate vaccination and screening programmes can prevent the development of cervical cancer. In clinical microbiology laboratories, PCR (Polymerase Chain Reaction) based tests are used to investigate the presence of HPV DNA in cervical swabs and for HPV genotyping. PCR requires specialised equipment such as a thermal cycler, a significant amount of time and trained personnel. Due to these requirements, the number of individuals who can access PCR tests is limited. As a solution to the known difficulties of PCR, a method known as Loop Mediated Isothermal Amplification (LAMP) has been developed. With the LAMP method, DNA amplification can be performed at a constant temperature of 60-65°C and in a short time without the need for complex laboratory equipment. In this study, we aimed to evaluate the efficiency of the LAMP method in the detection of HPV 16 and HPV 18 genotypes, which are responsible for the majority of cervical cancer cases, in clinical samples. Among the samples whose PCR tests were completed in the molecular microbiology laboratory for a year, HPV 16 and HPV 18 positive samples were collected prospectively In total, 259 HPV 16 and 29 HPV 18 were obtained from 288 cervical swab samples. After primer design for LAMP and LAMP optimisation, all isolates were included in LAMP experiments. PCR results were taken as reference to evaluate the performance of LAMP method in HPV 16 and HPV 18 detection. According to the results obtained, the sensitivity and specificity of the LAMP 7 method for HPV 16 were 97% and 100%, respectively, while the sensitivity and specificity for HPV 18 were 86% and 91%, respectively. The minimum amount of virus detected by the LAMP method was found to be 10 IU/mL for both HPV 16 and HPV 18 genotypes. The study showed that it was possible to detect HPV 16 and HPV 18 by LAMP method without DNA isolation from cervical swabs. It was observed that it was possible to obtain visual results by using Sybr Green stain or lateral flow strip in the detection of LAMP reaction products. As a result, it was concluded that the LAMP method can be a useful method for the detection of HPV 16 and HPV 18 in the laboratory or in bedside testing. It was concluded that studies are needed to improve the performance of the method by increasing the number and variety of samples.