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GBV-C, WNV & Dengue V in Ghanaian donors, women & children. C Li, D Candotti, JP Allain National Blood Service & University of Cambridge. Testing algorithm. Screening with Triplex NAT; probes labeled with: CY5 GBV-C, FAM WNV, HEX Dengue Confirmation with virus specific nested RT-PCR

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Gbv c wnv dengue v in ghanaian donors women children

GBV-C, WNV & Dengue V in Ghanaian donors, women & children

C Li, D Candotti, JP Allain

National Blood Service & University of Cambridge


Testing algorithm
Testing algorithm

  • Screening with Triplex NAT; probes labeled with:

  • CY5 GBV-C, FAM WNV, HEX Dengue

  • Confirmation with virus specific nested RT-PCR

  • Confirmed positive quantified by single virus QPCR for VL

  • Genotyping by sequencing E2 region and phylogenetic analysis

  • Antibody testing with Panbio kits


Quantification of gbv c rna load
Quantification of GBV-C RNA load

RNA from 200 l of plasma

Reference high + plasma: last + dilution defines arbitrary unit

GBV-C reference curve

40Ct = 1 AU/ml


Distribution of gbv c markers in blood donors and pregnant women li et al aids 2006 20 379 386
Distribution of GBV-C markers in blood donors and pregnant women (Li et al. AIDS 2006, 20:379-386)

Blood donations Pregnant women

HIV-1 status Neg Pos Neg Pos

N samples 140 96 140 86

% RNA+ 12.91 21.9 25 27.9

% anti-E2 + 3.8 14.5 18 17.3

% total markers 16.5 36.4 39.8 38.62

1. P<0.01 ; 2. Total is lower than RNA+Ab as some samples carry both


Gbv c wnv dengue v in ghanaian donors women children

35 women

30

25

20

% of total

15

10

5

0

>102

102

103

104

105

106

≥107

Distribution of HIV-1 RNA loads (IU/ml)

Distribution of HIV-1 RNA load according to GBV-C RNA status

GBV-C-

GBV-C+

(P< 0.01)


Gbv c wnv dengue v in ghanaian donors women children

GBV-C RNA positive women

(RNA load range, AU/ml)

HIV RNA status

Mothers

Cord blood

Positive

21

(1.1E3 - 5.6E5)

0

-

Negative

34

(1.7E0 - 5.6E5)

2 (5.9%)

-

pair 1458

pair 1473

1.2E5

3.7E5

2.6E3

2.8E1

Screening for GBV-C RNA in HIV-1 RNA positive and negative mothers and paired cord blood samples


Gbv c wnv dengue v in ghanaian donors women children

N= 157 women

44

91

125

105

71

68

18

16

14

12

10

% of total

RNA

IgG

8

6

4

2

0

<1

1-5

6-11

12-23

24-35

36-47

48-70

Age (months)

Prevalence of GBV-C markers with age



Gbv c wnv dengue v in ghanaian donors women children

50 women

40

% of total reactive

30

20

10

51-65y

6-11m

16-20y

21-25y

26-30y

31-35y

36-40y

41-45y

46-50y

12-23m

24-35m

36-47m

48-70m

Age distribution (months or years)

Age distribution of IgG antibody to West Nile Virus


Conclusions
Conclusions women

• GBV-C prevalence is high (16-40%) and genotype 1 is dominant

• GBV-C viraemia is twice as frequent as neutralising antibodies

• HIV-1 RNA load is lower in GBV-C viraemic patients

• GBV-C vertical transmission is rare and horizontal transmission appears predominant in West Africa (related to genotype?)

• Most WNV infections occur early in life, absence of viraemia, short window period --> minimal risk of transmission by transfusion

• Dengue virus does not seem prevalent in Ghana


Gbv c wnv dengue v in ghanaian donors women children

Acknowledgements women

KATH Blood Bank, Department of Medicine, Komfo Anokye Teaching Hospital, Kumasi, Ghana: Francis Sarkodie, Shirley Owusu-Ofori

INTS Paris, France: Jean-Jacques Lefrere, M Mariotti

Dept of Health & Human Services, F D A, USA: Indira Hewlett

Roche, USA: Jim Gallarda

National Institute for Biological Standards and Control, UK: Sally Baylis

University of Malaysia, Sarawak: Jane Cardosa