Simultaneous detection and genomic characterization of Zika virus Protein M, E and NS1 using optimized primers from Asian and African Lineage

Q3 Medicine

Vacunas Pub Date : 2024-01-01 DOI:10.1016/j.vacun.2023.07.003

Muhammed Mustapha Suleiman , Olatunji Matthew Kolawole

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引用次数: 0

Abstract

Purpose

Despite positive serological outcome, molecular confirmations have encountered little/no success either due to protocol accuracy, primer targets, or choice of sample amongst others. This study aims at providing an optimized protocol for the molecular detection of Zika virus amongst serologically positive respondent using samples from 2 selected hospitals in North Central Nigeria.

Materials and methods

About five (5) ml of blood samples was collected from a total of 400 participants for serological analysis, the IgM-positive samples were processed for molecular analysis using target primers from Asian and African lineage while a structured questionnaire was used to evaluate risk factors.

Results

Prevalence of 19% (38) and 45% (90) IgM and IgG positivity was recorded amongst respondent in Federal Medical Center (FMC), Keffi (R² = 1) while 36% (72) and 42% (84) was recorded in General Hospital (GH), Minna (R² = 1). The respective risk factors such as proximity of respondent to stagnant water or drainage channel, frequency of mosquito bite, prevention strategy, implementation of the prevention strategies for mosquito, and consumption of bushmeat were significant at set standard of P < 0.05. Molecular quantification revealed cut-off values (Ct) from 21.73 to 25.75 for all the 3 targeted protein while sequence analysis showed relatedness to deposited sequences in GenBank.

Conclusions

The abundance of the viral proteins as well as the genetic relatedness is indicative of presence of multiple strains of the virus or conservation of region across different geolocations. In lieu of the outcome, primers from multiple lineages is thereby recommended to forestall/overcome the challenge of cross-reactivity/false-negativity with Zika virus detection.

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使用来自亚洲和非洲血统的优化引物同时检测寨卡病毒蛋白 M、E 和 NS1 并确定其基因组特征

目的尽管血清学结果呈阳性，但由于方案准确性、引物目标或样本选择等原因，分子确证几乎没有成功。本研究旨在使用尼日利亚中北部两家选定医院的样本，为血清学阳性受访者中寨卡病毒的分子检测提供优化方案。材料和方法从总共 400 名参与者中采集约五（5）毫升血样进行血清学分析，使用亚洲和非洲血统的目标引物处理 IgM 阳性样本进行分子分析，同时使用结构化问卷评估风险因素。结果在凯菲联邦医疗中心（FMC）（R2 = 1）的受访者中，IgM 和 IgG 阳性率分别为 19%（38 人）和 45%（90 人），而在明纳综合医院（GH）（R2 = 1）的受访者中，IgM 和 IgG 阳性率分别为 36%（72 人）和 42%（84 人）。在 P < 0.05 的设定标准下，受访者与积水或排水沟的距离、蚊虫叮咬频率、预防策略、蚊虫预防策略的执行情况以及食用灌木肉等风险因素均具有显著性。分子定量分析显示，所有 3 种目标蛋白的临界值（Ct）从 21.73 到 25.75 不等，而序列分析表明与 GenBank 中保存的序列相关。因此，建议使用多系引物来防止/克服寨卡病毒检测中的交叉反应/假阴性问题。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Vacunas Medicine-Infectious Diseases

CiteScore

3.90

自引率

0.00%

发文量

138

审稿时长

62 days

期刊介绍： Sin duda una de las mejores publicaciones para conocer los avances en el campo de las vacunaciones preventivas, tanto en el ámbito de la investigación básica como aplicada y en la evaluación de programas de vacunaciones. Su alta calidad y utilidad la ha llevado a estar indexada en los prestigiosos índices IME y SCOPUS.