Table 1.

Molecular assays for RVFV.

Test	Test Type	Virus Detected	Target Gene	Biological Matrix	Reference Assay	Sample Size	Sensitivity	Specificity	Author
RVFV RT-nested PCR	Homemade one-step RT-PCR nested method	RVFV different strains (Gabek forest, Gordil, saint Floris, Arumwot, Belterra, ArD38661, AnD100286, MP 12)	NS coding region of S segment	Virus produced in Vero E6 cells; serum from infected mice	Virus isolation method	ND	0.5 pfu/reaction	Nd	Sall et al., 2001 [45]
RVFV quantitative real-time PCR	qRT-PCR with fluorescent signal from probes for quality control	RVFV; MP12, ZH501, ZH548, ArD38661, 74 HB59 strains	NS coding region of S segment	Virus produced in Vero E6 cells; serum from infected mice	ND	ND	50–100 copies/reaction	No amplification with Toscana, Icoraci, and Belterra closely related phlebovirus	Garcia et al., 2001 [46]
RT-Real-time PCR	5′ nuclease technology on a light cycler instrument	RVFV	G2 gene	Synthetic RNA	ND	ND	2835 geq/mL	no cross-reactivity with other HCV, HBV, HSV1, CMV, Modoc virus, Mycobacterium tuberculosis, Mycobacterium leprae, Borrelia spp., Leptospira spp., Neisseria spp., Plasmodium spp., Leishmania spp.	Drosten et al., 2002 [48]
RT-Real-time PCR homemade	Fluorescent nested PCR TaqMan assay	RVFV	S segment	Synthetic RNA	ND	ND	100 copies/reaction	SFNV cell culture	Weidmann et al., 2008 [47]
Real-time qRT-PCR homemade	qRT-PCR with fluorescent reporter dye detected at each PCR cycle	RVFV	G2 gene	Plasma of suspected patients with HVF	272 RVFV confirmed cases	2ND	100 infectious particles/mL	IgM anti RVFV positive sera 100%	Njenga et al., 2009 [49]
RT-LAMP homemade	Reverse transcription-loop-mediated isothermal amplification with a vertical	RVFV	L segment	Serum samples	TaqMan Real Time	64	Whole blood: LLOD: 10 copies RNA/reaction	No cross reactivity with phleboviruses; flaviviruses and chikungunja virus	Peyrefitte et al., 2008 [50]
RT-LAMP homemade	Reverse transcription-loop-mediated isothermal amplification with a vertical	RVFV	L segment	Bleed samples from sheep (n = 20), human plasma from suspected cases (n = 65); 3 liver, kidney, serum from animals			Whole blood: LLOD: 10 copies/ reaction	Six African phleboviruses and unrelated arbovirus did not give cross reactivity	Le Roux et al., 2009 [51]
RT-LAMP homemade	Reverse transcription-loop-mediated isothermal amplification	RVFV	S segment	Synthetic RNA	Real-time RT-PCR	ND	whole blood: LLOD: 1.94 copies/microliters within 60 min	No cross reactivity with JEV, H3N2 influenza virus, EBOV, MARV	Han et al., 2020 [52]
RT-LAMP homemade	Reverse transcription-loop-mediated isothermal amplification	RVFV	M segment	Blood samples	Real-time RT-PCR	130	98.36% sensitivity	100%; no cross-reactivity with PPR and capripox viruses	Wekesa et al., 2023 [54]
Isothermal recombinase polymerase amplification (RPA)	Isothermal exponential nucleic acid amplification and detection method	RVFV	S segment	Synthetic RNA	ND	ND	19RNA molecules/reaction	No cross reactivity with Yersinia pestis, Francisella tularensis, Bacillus antracis, vaccinia virus, Ebola virus, Marburg virus, Crimean–Congo virus and phleboviruses	Euler et al., 2012 [53]
RT-qPCR genotyping assay	One step RT-qPCR for typing different strains of RVFV, melting curve to identify different strains of RVFV	RVFV	L, M, S segments	ND	Sanger sequencing		ND		Balaraman et al., 2023 [56]
BioT DNA multiplex PCR-enzyme hybridization assay	Multiplex RT-PCR	RVFV	GP2 gene	196 swabs, 45 skin swabs,15 serum, 7 sputum	ND	260 clinical samples	105–106 copies/mL with nucleic acid extraction	No cross reactivity with Influenza A, EBV, CMV, RSV A, ADV C, human metapneumovirus	He et al., 2009 [55]
Oligonucleotide microarray	Microarray	RVFV	GP gene	Culture samples	Real-time PCR	60	100%		Yao et al., 2021 [57]
Real-time qRT-PCR commercial	qRT-PCR with fluorescent reporter dye detected at each PCR cycle	RVFV					0.89 copies/μL	cross-reactivity with flavivirus, Marburg virus, and Ebola virus	[61]

ADV, adenovirus; CMV, cytomegalovirus; RSV, respiratory syncytial virus; JEV, Japanese encephalitis virus.