Utilization of 18s ribosomal RNA LAMP for detecting Plasmodium falciparum in microscopy and rapid diagnostic test negative patients

• Published in: PloS one 2022-10, Vol.17 (10), p.e0275052
• Main Authors: Aninagyei, Enoch, Boakye, Adjoa Agyemang, Tettey, Clement Okraku, Ntiri, Kofi Adjei, Ofori, Samuel Ohene, Tetteh, Comfort Dede, Aphour, Thelma Teley, Rufai, Tanko
• Format: Article
• Language: English
• Subjects: Biology and Life Sciences; Child; Data analysis; Demographic variables; Deoxyribonucleic acid; Diagnosis; Diagnostic systems; Diagnostic tests; Diagnostic Tests, Routine - methods; Diarrhea; DNA; DNA polymerase; Female; Gene amplification; Health risks; Health services; Herpes labialis; Humans; Identification and classification; Laboratories; Malaria; Malaria - diagnosis; Malaria, Falciparum - diagnosis; Male; Males; Medical diagnosis; Medicine and Health Sciences; Microscopy; Molecular Diagnostic Techniques; Nucleic Acid Amplification Techniques - methods; Parasites; Patients; Plasmodium; Plasmodium - genetics; Plasmodium falciparum; Plasmodium falciparum - genetics; Reagent Kits, Diagnostic; Research and Analysis Methods; Ribosomal RNA; RNA, Ribosomal, 18S - genetics; rRNA 18S; Sensitivity and Specificity; Vector-borne diseases; Vomiting
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0275052

In this study, Plasmodium falciparum was detected in patients that were declared negative for malaria microscopy and rapid diagnostic test kit (mRDT), using Plasmodium 18s rRNA loop-mediated isothermal amplification (LAMP) technique. The main aim of this study was to assess the usefulness of LAMP assay for detecting pre-clinical malaria, when microscopy and mRDT were less sensitive. DNA was obtained from 100 μL of whole blood using the boil and spin method. Subsequently, the Plasmodium 18s rRNA LAMP assay was performed to amplify the specific Plasmodium 18s rRNA gene. Microscopy and mRDT negative samples [697/2223 (31.2%)] were used for this study. Compared to frequencies obtained for the other demographic variables, most of the patients were < 6 years (37.7%), females (59.0%), peri-urban dwellers (39.0%) and patients that sought outpatient department services (39.3%). Overall, the prevalence of Plasmodium 18s rRNA was 17.5%. when stratified by study variables, Plasmodium 18s rRNA LAMP positivity was higher in patients over 30 years [58/122 (54.2%)], males [69/122 (56.5%)], rural dwellers [69/122 (56.5%)] and patients that sought OPD services [68/122 (55.7%)]. The risk of being infected with Plasmodium when routine tests were negative was higher in 15-30-year group (OR = 3.03, 95% CI: 1.6-5.8, p = 0.0007), patients > 30 years (OR = 15.2, 95% CI: 8.3-27.7, p