Developments in the detection and understanding of meningococcal carriage, and implications for studies of the impact of MenB vaccines https://www.meningitis.org/mrf-conference-2017

1. 15/11/2017
1
Developments in the detection and
understanding of meningococcal
carriage and implications for
studies of the impact of MenB
vaccines
@adamhfinn
MRF London 15th Nov 2017
tinyurl.com/adammrf2017
MenC changing
schedules
Birth 10 years 20 years
ACYW
tinyurl.com/adammrf2017
MenW cases in England,
2005/06-2014/15
tinyurl.com/adammrf2017
Cortesia de
S
Ladhani,
PHE,
RU Cortesia de
S
Ladhani,
PHE,
RU
2014/15
2015/16
2016/17
UK cumulative incidence of MenB
cases by age group 2011-2017
tinyurl.com/adammrf2017
Secondary endpoints
4CMenB vaccine
MenB Controls Efficacy
95% CI
All B 233/2489
9.4%
262/2576
10.2%
+15.6%
-­11.0 to +35.9%
All Nm 797/2489
32.0%
885/2576
34.4%
+18.2%
+3.4 to +30.8%
B C W Y 449/2489
18.0%
539/2576
20.9%
+26.6%
+10.5 to +39.9%
Read et al., ESPID 2013, Lancettinyurl.com/adammrf2017
Carriage studies
• Pfizer study – freezing and batching
• Portugal 1 – PCR and cx-amp PCR
• BrisMenNHC - longitudinal
– (MRF1) virology; GSK genogrouping
• Portugal 2 – saliva
• SPIT (MRF2&3) – lots more saliva
• Better understanding of biology
• Better sampling and lab methods
tinyurl.com/adammrf2017

2. 15/11/2017
2
Throat Swabs / OPS
Direct plating
Stored STGG broth
qPCR
tinyurl.com/adammrf2017
Meningococcal carriage
density in 87 young adults
0
5
10
15
20
25
30
35
40
45
50
0 to <10 10 to <100 100 to <1000 1000 to <10000
CFU/ml
Number of students
tinyurl.com/adammrf2017
Mening’ carriage density
Finn et al PIDJ 2016
Culture – left/hatched
qPCR – right/solid
Upper grey sections non-­concordant results
tinyurl.com/adammrf2017
tinyurl.com/adammrf20
17
Oxford
Bristol V1 V2 V3 V4 V5 V6
Other
centres
V
V
15-­19 year olds
Into 1.5ml STGG
broth on site
Meningococci
determined by
q-­PCR for sodC
Xfer to lab within
2-­6 hours
frozen -­80 °C
Swab and questionnaire at each time point
£5 thank you for their time for V2-­6
BrisMenNHC
Density over time within
individuals appears highly
variable
tinyurl.com/adammrf20
17
0
1000
2000
3000
4000
5000
6000
Gene copies/ml
Visit date
Density over time within
individuals appears highly
variable
tinyurl.com/adammrf20
17
0
1000
2000
3000
4000
5000
Gene copies/ml
Visit date

3. 15/11/2017
3
Density over time within
individuals appears highly
variable
tinyurl.com/adammrf20
17
0
200
400
600
800
1000
1200
Gene copies/ml
Visit date
Density over time within
individuals appears highly
variable
tinyurl.com/adammrf20
17
0
500
1000
1500
2000
2500
Gene copies/ml
Visit date
Density over time within
individuals appears highly
variable
tinyurl.com/adammrf20
17
0
2000
4000
6000
8000
10000
12000
14000
16000
18000
Gene copies/ml
Visit date
tinyurl.com/adammrf20
17
Characteristic Group coeff 95% CI p
Gender Female* 0 (reference) 0.020
Male 0.40 0.06 0.73
Age group 15/16 0.15 -­0.24 0.54 0.218
17* 0 (reference)
18 -­0.38 -­0.80 0.03
19 0.38 -­1.12 1.88
School year Year 12* 0 (reference) 0.006
Year 13 0.70 0.27 1.14
Other 0.18 -­1.77 2.14
Ethnicity White* 0 (reference) 0.033
Asian -­0.61 -­1.27 0.04
Black 0.50 -­0.31 1.32
Mixed/other -­0.76 -­1.53 0.01
Antibiotic use None* 0 (reference) 0.073
Stopped in last month0.67 -­0.04 1.39
Stopped in last week 0.15 -­0.79 1.08
Currently using -­0.68 -­1.41 0.05
Pub days in week 0* 0 (reference) 0.068
1 0.10 -­0.19 0.38
2–7 0.54 0.08 0.99
Has smoker GF/BF No* 0 (reference) 0.076
Yes -­0.94 -­1.98 0.10
Month of year Jan* 0.00 0.00 0.00 0.193
Feb 0.02 -­0.37 0.40
Mar -­0.12 -­0.51 0.27
Apr 0.00 -­0.44 0.44
May -­0.37 -­0.88 0.14
Sep -­0.70 -­1.34 -­0.06
Oct -­0.16 -­0.64 0.32
Nov 0.15 -­0.27 0.58
Dec 0.17 -­0.34 0.68
Constant -­1.33 -­1.79 -­0.86 0.000
We found
• Gender
• School year
• Ethnicity
• Antibiotic use
• Pub visits
• Partner who
smokes
were associated
with Nm density
Density vs. month
tinyurl.com/adammrf2017
Associations between
Meningococcal Carriage &:
P=0.0564
P=0.0354
N.
meningitidis+
N.
meningitidis-
Total
Virus+ 14 105 119
Virus- 88 1173 1261
Total 102 1278 1380
RhV+ 12 81 93
RhV- 90 1197 1287
Total 102 1278 1380
Rhinovirus
Presence
Virus
Presence
N.B. Percentages as a proportion of total Viral/RhV +/-
12.9% 87.1% 100%
7.0% 93.0% 100%
11.8% 88.2% 100%
7.0% 93.0% 100%
tinyurl.com/adammrf2017

4. 15/11/2017
4
Results-By Calendar Month
0
2
4
6
8
1 0
1 2
1 5
2 0
2 5
M o n th
Percentage(%)
F lu A
R S V
R h V
N . m e n in g it id is
S e p O ct N o v D e c J a n F e b M a r A p r
2 30 2 30 2 200 1 38 2 56 2 46 60n =
tinyurl.com/adammrf2017
See Liz
Oliver’s
poster here
Comparing methods
tinyurl.com/adammrf2017
1
45
87
2442
121
1493 46
66
1614
Culture
PCR
Culture amplified
PCR
Cx 7.6%
PCR 8.8%
Cx+PCR 11.4%
CaPCR 15.9%
Cx+PCR+CaPCR 18.3%
Multiple genogroups
Ø 5 of the 1,813 samples detect multiple genogroups in one sample run of qPCR
and the relevant Ct values
Ø Ct £ 36 – positive for Nm generic sodC and genogroups B, C, W, X and Y.
tinyurl.com/adammrf2017
See Kirsty
Nelson’s
poster here
Saliva
tinyurl.com/adammrf2017
Increasing sensitivity…
tinyurl.com/adammrf2017
N=1005 18-­25 year old medical/pharmacy students
Coimbra, Portugal
Increasing sensitivity…
tinyurl.com/adammrf2017

5. 15/11/2017
5
All meningococci - PCR
tinyurl.com/adammrf2017
Throat swabsSaliva
Direct
Culture-­amp’d
126
72
54
71
12.5%
19.6%
33.1%
136 34
33
5
20
23
11
52
14
5
36.1%
119
17
30
25
8
11
12
1419
6
GpB meningococci - PCR
tinyurl.com/adammrf2017
Throat swabsSaliva
Direct
Culture-­amp’d
PCR 1.7%
+caPCR 3.1%
+Sal 3.3%
2
0
8
3
12
14
1+caPCR 3.6%
1
1
3
0
0
1
3
0
1
Gps BWXY meningococci - PCR
tinyurl.com/adammrf2017
Throat swabsSaliva
Direct
Culture-­amp’d
PCR 2.2%
+caPCR 4.4%
+Sal 5.0%
4
0
10
3
19
14
1+caPCR 5.5%
4
2
5
0
1
1
3
0
2
SPIT
• 433 16-18 year olds – 419/397/296 V1-3
– Approx. 50% participation
• All offered 2 doses Bexsero
• OPS at vaccine visits & 3 months later
– 71% of V1s came for final swab
• Weekly salivas from pre-vaccine to end
• MRF have funded direct sodC PCR from
all OPS and salivas. Further funding needed
• Questionnaires on demographics and
motivation to take part…see Nicola Kelly’s
poster heretinyurl.com/adammrf2017
SPIT study
OPS/Saliva samples
tinyurl.com/adammrf2017
NanoString nCounter system
Gene detection &
counting nCounter
tinyurl.com/adammrf2017

6. 15/11/2017
6
Conclusions
• Meningococcal carriage density is
dynamic – possibly driven by viral
infections
• Overall crude carriage rates in
vaccine recipients may be relatively
poor predictors of vaccine impact on
transmission
• Viable meningococci can be found in
saliva – implications for transmission
and precautions
tinyurl.com/adammrf2017
Conclusions
• Significant sampling and laboratory
methodological advances now available
to make carriage studies more sensitive
and informative.
• IMD not that rare – spend money on
vaccines not lottery tickets!
• Thanks to MRF for support
tinyurl.com/adammrf2017