Department of Biomedical Sciences, School of Basic and Biomedical Sciences, University of Health and Allied Sciences, Ho, Volta Region, Ghana.
Ghana Health Service, Greater Accra Region, Mayera-Faase Health Centre, Mayera-Faase, Ghana.
PLoS One. 2022 Oct 6;17(10):e0275052. doi: 10.1371/journal.pone.0275052. eCollection 2022.
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<0.001), males (OR = 2.1, 95% CI: 1.4-3.2, p = 0.0002) and rural dwellers (OR = 2.2, 95% CI:1.4-3.6, p = 0.0009). However, risk was lower in post-natal children (OR = 0.3, 95% CI: 0.18-0.51, p<0.001). Majority (81.5%) of the infected patients presented with headache, herpes labialis, diarrhea and vomiting. We demonstrated the lack of sensitivities of microscopy and mRDT for one-time diagnosis of malaria. Therefore, it is essential to utilize a sensitive technique such as Plasmodium 18s rRNA LAMP to increase the detection rate of Plasmodium infection.
在这项研究中，使用疟原虫 18s rRNA 环介导等温扩增（LAMP）技术，在宣布疟疾显微镜检查和快速诊断检测试剂盒（mRDT）阴性的患者中检测到恶性疟原虫。本研究的主要目的是评估当显微镜检查和 mRDT 不太敏感时，LAMP 检测对检测临床前疟疾的有用性。使用煮沸和旋转法从 100 μL 全血中获得 DNA。随后，进行疟原虫 18s rRNA LAMP 检测以扩增特定的疟原虫 18s rRNA 基因。对 [697/2223（31.2%）] 个显微镜和 mRDT 阴性样本进行了此项研究。与其他人口统计学变量的频率相比，大多数患者年龄<6 岁（37.7%）、女性（59.0%）、城乡结合部居民（39.0%）和门诊患者（39.3%）。总体而言，疟原虫 18s rRNA 的流行率为 17.5%。按研究变量分层时，30 岁以上患者 [58/122（54.2%）]、男性 [69/122（56.5%）]、农村居民 [69/122（56.5%）] 和门诊患者 [68/122（55.7%）] 中疟原虫 18s rRNA LAMP 阳性率较高。在常规检测为阴性时，15-30 岁组感染疟原虫的风险更高（OR=3.03，95%CI：1.6-5.8，p=0.0007），30 岁以上患者（OR=15.2，95%CI：8.3-27.7，p<0.001）、男性（OR=2.1，95%CI：1.4-3.2，p=0.0002）和农村居民（OR=2.2，95%CI：1.4-3.6，p=0.0009）。然而，产后儿童的风险较低（OR=0.3，95%CI：0.18-0.51，p<0.001）。大多数（81.5%）感染患者出现头痛、唇疱疹、腹泻和呕吐。我们证明了显微镜检查和 mRDT 单次诊断疟疾的敏感性不足。因此，有必要利用疟原虫 18s rRNA LAMP 等敏感技术来提高疟原虫感染的检测率。
