Table 1 Characteristics of the included studies

1.

Asua et al. [2]

Uganda

Malaria positive (499)

6 months to 10 years

Dried blood spots

Chelex extraction kits

18S rRNA

Nested PCR

Pf/Pm (19)

Pf/Po (14)

Pf/Pv (3)

Pm/Pv (1)

Total = 37

Pf/Pm/Po (1)

2.

Barber et al. [3]

Malaysia (2009–2011)

Malaria positive (653)

P. knowlesi (IQR 20–50 years),

P. falciparum (IQR 9–31 years),

P. vivax

(19 years, IQR 7–32 years)

Triple infection: < 5 (1), 10–14 (1), 30–34 (1), 80–84 (1)

Dual infection: < 5 (1), 5–9 (2), 10–14 (5), 15–19 (3), 20–24 (2), 25–29 (2), 30–34 (4), 35–39 (3), 40–44 (5), 45–49 (2), 50–54 (2), 70–74 (2), 75–79 (1)

NA

NA

18S rRNA

Nested PCR

Pv/Pk (36)

Pf/Pk (6)

Pm/Pk (1)

Pf/Pm (1)

Total = 44

Pf/Pv/Pk (2)

Pv/Pm/Pk (2)

3.

Calderaro et al. [4]

Italy (2000–2007)

Febrile patients (701)

NA

EDTA blood

High Pure Template Preparation kit (Roche

Diagnostics, Mannheim, Germany)

18S rRNA

Nested PCR

Pf/Po (3)

Pf/Pm (1)

Total = 4

Pf/Pm/Po (2)

4.

Camargo-Ayala et al. [5]

Colombia (2012–2015)

Febrile patients (671)

< 5, 5–18, 18–60, and > 60 years

Triple infection: < 5 (4), 5–18 (2), 18–60 (6), > 60 (2)

Dual infection: < 5 (16), 5–18 (56), 18–60 (139), > 60 (15)

EDTA blood

Pure Link Genomic DNA mini kit (Invitrogen)

18S rRNA

Nested PCR

Pv/Pm (190)

Pf/Pv (25)

Pf/Pm (10)

Total = 225

Pf/Pm/Pv (14)

5.

Dhangadamajhi et al. [6]

India (2008)

Febrile and asymptomatic (242)

0–5 years, 6–15, and > 15 years

Triple infection: 0–5 (1), 6–15 (2), > 15 (2)

Dual infection: 0–5 (7), 6–15 (32), > 15 (39)

EDTA blood

Phenol–Chloroform extraction

18S rRNA

Nested PCR

Pf/Pv (15)

Pf/Pm (54)

Pv/Pm (10)

Total = 79

Pf/Pm/Pv (5)

6.

Dormond et al. [40]

Switzerland (2004–2008)

Malaria positive (89)

>16 years

EDTA blood

MagNA Pure LC DNA isolation kit (Roche, Basel, Switzerland)

18S rRNA

Nested PCR

Pf/Po (3)

Pf/Pm (2)

Total = 5

Pf/Pm/Po (1)

7.

Fuehrer et al. [41]

Bangladesh:

Chittagong Hill Tracts (2007–2008)

Malaria Research Initiative

Bandarban field site (2008–2009)

Febrile patients (379)

Any age

Dried blood spots

Modified Chelex-based technique, the InstaGene Whole Blood

Kit (Bio-Rad Laboratories, Hercules, CA)

18S rRNA

Nested PCR

Pf/Pv (21)

Pf/Pm (2)

Total = 23

Pf/Pm/Pv (2)

8.

Gabrielli et al. [42]

Congo (2014)

Residents

(306)

1 week to < 5 years

Dried blood spots

The commercial

kit Dried Blood spot DNA isolation Kit (Norgen Biotek

Corp, Ontario, Canada)

18S rRNA

Nested PCR

Pf/Pm (13)

Pf/Po (2)

Pf/Pv (1)

Total = 16

Pf/Pm/Pv (1)

9.

Hopkins et al. [22]

Uganda (2010–2011)

Residents (272)

5–81 years: 5–10, 11–20, and ≥ 21 years

EDTA blood

Phenol–Chloroform extraction

18S rRNA

Nested PCR and qPCR

Pf/Pm (38)

Pf/Po (8)

Total = 46

Pf/Pm/Po (10)

10.

Jiang et al. [43]

Myanmar (2008)

Malaria positive (146)

NA

Dried blood spots

Chelex extraction kits

18S rRNA

Nested PCR

Pf/Pk (13)

Pv/Pk (13)

Pf/Pv (10)

Total = 36

Pf/Pv/Pk (2)

11.

Kasehagen et al. [11]

Papua New Guinea (2001–2003)

Residents (16,209)

< 2, 2.0–3.9, 4.0–6.9, 7.0–9.9, 10.0–19.9, 20.0–39.9, and ≥ 40 years

EDTA blood

QIAamp 96 DNA Blood Kit (QIAGEN, Valencia, CA)

18S rRNA

LDR-FMA

Pf/Pv (72)

Pf/Pm (32)

Pf/Po (13)

Pv/Pm (17)

Pv/Po (9)

Pm/Po (4)

Total = 147

Pf/Pm/Pv (28)

Pf/Pv/Po (5)

Pf/Pm/Po (4)

Pv/Pm/Po (4)

Pf/Pv/Pm/Po (11)

12.

Krishna et al. [18]

India (2015)

Doubtful microscopic (355)

≤ 1, > 1–4, >4–8, > 8–14, and > 14 years

Dried blood spots

FavorPrep Genomic DNA Mini Kit (Favorgen Biotech Corp., Taiwan).

18S rRNA

Nested PCR

Pf/Pv (59)

Pf/Pm (3)

Total = 62

Pf/Pm/Pv (5)

Pf/Pv/Po (1)

Pf/Pv/Pm/Po (1)

13.

Lorenzetti et al. [44]

Brazil (2003–2005)

Malaria positive (115)

18–52 years

EDTA blood

Phenol–Chloroform extraction

18S rRNA

Nested PCR

Pf/Pm (2)

Pf/Pv (28)

Total = 30

Pf/Pm/Pv (1)

14.

Marques et al. [45]

Mozambique (2001–2002)

Residents (308)

1–82 years

EDTA blood

Phenol–Chloroform extraction

18S rRNA

Nested PCR

Pf/Pm (70)

Pf/Po (10)

Total = 80

Pf/Pm/Po (9)

15.

May et al. [48]

Nigeria (1996–1997)

Residents (593)

Children in Abanla (1–11 years), Children in Ibadan (3–8 years), Children in health centres in Ibadan (0.8–11 years), Healthy adults (15–56 years)

EDTA blood

DNA-Easy Kit (Invitrogen, St. Louis, MO)

18S rRNA

Nested PCR

Pf/Pm (41)

Pf/Po (12)

Total = 53

Pf/Pm/Po (27)

16.

Mehlotra et al. [13]

Papua New Guinea (1998–1999)

Residents (1848)

All groups < 1–85 years: 2–4, 5–9, ≥ 10 years

Triple infection: 2–4 (2), 5–9 (8), ≥ 10 (24)

Dual infection: 2–4 (14), 5–9 (51), ≥ 10 (115)

EDTA blood

QIAamp 96 spin blood kits

(QIAGEN, Valencia, CA)

18S rRNA

Nested PCR

Pf/Pv (116)

Pf/Pm (27)

Pf/Po (11)

Pv/Pm (21)

Pv/Po (4)

Pm/Po (3)

Total = 182

Pf/Pm/Pv (23)

Pf/Pv/Po (9)

Pf/Pm/Po (1)

Pf/Pv/Pm/Po (3)

17.

Mehlotra et al. [12]

Papua New Guinea (1996)

Malaria positive (173)

5–10 and > 11 years

Triple infection: 5–10 (1), > 11 (0)

Dual infection: 5–10 (6), > 11 (9)

EDTA blood

QIAamp 96 or individual spin blood

kits (QIAGEN, Valencia, CA)

18S rRNA

Nested PCR

Pf/Pv (40)

Pf/Pm (16)

Pf/Po (4)

Pv/Pm (3)

Total = 63

Pf/Pm/Pv (27)

Pf/Pv/Po (8)

Pf/Pm/Po (5)

Pv/Pm/Po (1)

Pf/Pv/Pm/Po (9)

18.

Mueller et al. [14]

Papua New Guinea (2005)

Residents (2527)

< 10 and ≥ 20 years

EDTA blood

QIAmp 96 DNA Blood kits (Qiagen, CA)

18S rRNA

A semi-quantitative post-PCR, ligase detection reaction/fluorescent microsphere assay (LDR-FMA)

Pf/Pv (363)

Pf/Pm (136)

Pf/Po (27)

Pv/Pm (26)

Pv/Po (7)

Pm/Po (1)

Total = 560

Pf/Pm/Pv (99)

Pf/Pv/Po (33)

Pf/Pm/Po (8)

Pv/Pm/Po (6)

Pf/Pv/Pm/Po (17)

19.

Nino et al. [46]

Colombia (2015–2016)

Febrile patients (1392)

NA

EDTA blood

Pure Link Genomic DNA mini kit (Invitrogen)

18S rRNA

Nested PCR

Pf/Pv (111)

Pv/Pm (340)

Pf/Pm (29)

Total = 480

Pf/Pm/Pv (52)

20.

Pati et al. [19]

India

Febrile patients (1589)

Severe malaria (15–65 years)

Dried blood spots

Chelex extraction kits

18S rRNA

Nested PCR

Pf/Pv (11)

Pf/Pm (4)

Pv/Pm (3)

Total = 18

Pf/Pm/Pv (5)

21.

Perandin et al. [20]

Italy

Febrile patients (122)

NA

EDTA blood

The High Pure PCR template preparation kit (Roche, Indianapolis,

Ind.)

18S rRNA

Nested PCR

Pf/Po (1)

Pf/Pm/Po (1)

22.

Peruzzi et al. [21]

Italy (2005–2006)

Febrile patients (139)

2–49 years

Triple infection: 19 years

EDTA blood

High Pure PCR Template

Preparation Kit” (Roche)

18S rRNA

Nested PCR

None

Pf/Pm/Po (1)

23.

Pongvongsa et al. [28]

Laos-Vietnam border (2010)

Residents (3059)

Sample for PCR (135)

P. knowlesi (2–15 years)

Triple infection (Pf/Pv/Pk): mean 6.43 years (7)

Dual infection (Pk/Pv): mean 7.4 years (5)

Dried blood spots

QIAamp DNA micro kit

(QIAGEN, Tokyo, Japan)

18S rRNA

Nested PCR

Pf/Pv (15)

Pf/Pm (2

Pv/Pk (5)

Total = 22

Pf/Pm/Pv (1)

Pf/Pv/Pk (7)

24.

Putaporntip et al. [25]

Thailand (2006–2007)

Febrile patients (1874)

1–81 years

Dried blood spots

Qiagen DNA Mini Kit

18S rRNA

Nested PCR

Pf/Pv (200)

Pf/Pm (6)

Pf/Po (2)

Pf/Pk (5)

Pv/Pm (8)

Pv/Po (4)

Pv/Pk (4)

Pm/Po (1)

Total = 230

Pf/Pm/Pv (4)

Pf/Pv/Po (7)

Pf/Pv/Pm/Po (1)

25.

Rubio et al. [38]

Spain (1997–1998)

Febrile patients (168)

NA

EDTA blood

Modified Chelex-based technique, the InstaGene Whole Blood

Kit (Bio-Rad Laboratories, Hercules, CA)

18S rRNA

Nested PCR

Pf/Pm (4)

Pf/Po (1)

Pf/Pv (3)

Total = 8

Pf/Pm/Pv (1)

26.

Rubio et al. [39]

Equatorial Guinea (1996)

Febrile patients (159)

< 6 years

Dried blood spots

5% Chelext-100 Resin (Bio-Rad Laboratories,

Hercules, CA)

18S rRNA

Nested PCR

Pf/Pm (36)

Pf/Po (3)

Pf/Pv (2)

Total = 41

Pf/Pv/Po (3)

27.

Sitali et al. [29]

Zambia (2012)

Residents (873)

< 6 years

Dried blood spots

Chelex extraction kits

18S rRNA

Nested PCR

Pf/Pm (31)

Pf/Po (10)

Pf/Pv (1)

Total = 42

Pf/Pm/Po (6)

Pf/Pm/Pv (1)

28.

Sluydts et al. [15]

Cambodia (2012)

Residents (5793)

2–5, 5–14, 15–39, and ≥ 40 years

Dried blood spots

Instagene^® Matrix resin (Bio-Rad, Singapore)

18S rRNA

Two-step Real-time PCR

Pf/Pv (56)

Pf/Pm (5)

Pf/Po (1)

Pv/Pm (3)

Pv/Po (11)

Total = 76

Pf/Pm/Pv (4)

Pf/Pv/Po (1)

Pf/Pv/Pm/Po (1)

29.

Steenkeste et al. [16]

Cambodia (2001)

Residents (337)

NA

Dried blood spots

the Instagene resin (Bio-Rad,

USA)

18S rRNA

Nested PCR

Pf/Pv (52)

Pf/Pm (15)

Pf/Po (4)

Total = 71

Pf/Pv/Po (7)

Pf/Pm/Pv (24)

Pf/Pv/Pm/Po (8)

30.

Steenkeste et al. [17]

Cambodia (2001)

Residents (134)

< 1– > 60 years: < 1, 1, 2, 3, 4, 5–9, 10–14, 15–19, 20–39, 40–60, and > 60 years

Dried blood spots

Instagene resin (Bio-Rad, Germany) a

18S rRNA

Nested PCR

Pf/Pm (4)

Pf/Po (2)

Pf/Pv (18)

Pv/Pm (3)

Total = 27

Pf/Pm/Pv (7)

Pf/Pv/Po (3)

Pf/Pv/Pm/Po (3)

31.

Subissi et al. [23]

Uganda (2010)

Residents (509)

1–5, 6–10, and > 20 years

Triple infection: 1–5 (5), 6–10 (5), > 20 (0)

Dual infection: 1–5 (27), 6–10 (24), > 20 (2)

Dried blood spots

QIAamp mini kit (QIAGEN, Venlo, The Netherlands)

18S rRNA

Nested PCR

Pf/Pm (39)

Pf/Po (14)

Total = 53

Pf//Pm/Po (10)

32.

Toma et al. [27]

Laos (1997)

Residents (336)

< 11– > 50 years: 2–68 years in Phavang and 0–75 years in Sisomsouen

EDTA blood

GFX Genomic Blood DNA Purification

Kit (Pharmacia Biotech).

18S rRNA

Nested PCR

Pf/Pv (19)

Pf/Pm (2)

Pv/Pm (2)

Pv/Po (1)

Total = 24

Pf/Pm/Pv (1)

Pf/Pv/Po (1)

Pf/Pv/Pm/Po (1)

33.

Woldearegai et al. [47]

Gabon (2016)

Residents (834)

1–96 years: 1–5, 6–10, 11–15, 16–20, 21–25, 26–30, 31–40, 41–50, 51–60, 61–70, 71–80, and 81–96 years

EDTA blood

QIAsymphony DSP DNA kit

18S rRNA

Nested PCR

Pf/Pm (123)

Pf/Po (43)

Pm/Po (1)

Total = 167

Pf//Pm/Po (51)

34.

Zhou et al. [24]

Thailand (1995–1996)

Residents (548)

NA

Thick smear

30% Chelex-100

18S rRNA

Nested PCR

Pf/Pv (10)

Pf/Pm (6)

Pv/Pm (19)

Pv/Po (1)

Total = 36

Pf/Pm/Pv (49)

Pf/Pv/Po (2)

Pv/Pm/Po (1)

Pf/Pv/Pm/Po (7)

35.

Zhou et al. [26]

China (2008–2012)

Febrile patients (562)

NA

Dried blood spots

QIAamp

DNA Mini Kit (QIAGEN China (Shanghai)

18S rRNA

Nested PCR

Pf/Pv (67)

Pv/Po (2)

Pf/Pk (2)

Pf/Pm (2)

Po/Pm (1)

Total = 74

Pf/Pv/Po (3)

Pf/Pm/Pv (1)

Pv/Pm/Po (1)

Pf/Pv/Pm/Po (1)

Total = 12,023

2 species = 3059

Pf/Pv = 1318

Pf/Po = 188

Pf/Pm = 775

Pf/Pk = 26

Pv/Pm = 645

Pv/Po = 39

Pv/Pk = 58

Pm/Po = 10

Pm/Pk = 1

3 species = 601

Pf/Pm/Pv = 355

Pf/Pv/Po = 83

Pf/Pm/Po = 137

Pf/Pv/Pk = 11

Pv/Pm/Po = 13

Pv/Pm/Pk = 2

4 species = 53