Salah A. Ibrahim

Combination of reverse transcriptase PCR analysis and immunoglobulin M detection on filter paper blood samples allows diagnostic and epidemiological studies of measles.

ABSTRACT As measles control and elimination campaigns progress, laboratory confirmation of clinically diagnosed measles cases becomes increasingly important. However, in many tropical countries collection and storage of clinical specimens for this purpose are logistically complicated. In this study it is shown that blood samples spotted on filter paper are suitable for the laboratory diagnosis of measles using a combination of reverse transcriptase PCR (RT-PCR) analysis and immunoglobulin M (IgM) detection. First, it was shown that in vitro measles virus (MV)-infected cells diluted in human blood and spotted on filter paper can be detected by RT-PCR. Small amounts of infected cells remained detectable after 25 weeks of storage of the filter paper at room temperature, 4 weeks at 37°C, or 2 weeks at 45°C. Subsequently, this RT-PCR was applied to filter paper blood samples collected from 117 clinically diagnosed measles patients in Sudan in 1997 and 1998. Prior laboratory diagnosis had confirmed 90 cases as acute MV infections, while 27 proved to be nonmeasles rash disease cases. Positive RT-PCR signals were detected in filter paper blood samples of 43 of the 90 confirmed cases (48%) but in none of the 27 nonmeasles cases. In addition, MV-specific IgM levels measured in reconstituted filter paper samples correlated well with those measured in plasma samples. Measles diagnosis based on the combination of filter paper RT-PCR and IgM detection had a sensitivity and specificity of 99 and 96%, respectively. An advantage of this diagnostic approach is that sequencing of RT-PCR products allows phylogenetic analysis of the MV strain involved.

Neonatal resuscitation courses for pediatric residents: comparison between Khartoum (Sudan) and Padova (Italy)

Background:  The efficacy of a Neonatal Resuscitation Program (NRP) has been previously evaluated in developed countries, but there is a lack of information regarding the impact of this teaching program in developing countries. Our aim was to compare the knowledge gained by University of Khartoum (Sudan) and University of Padova (Italy) pediatric residents following participation in the NRP course.

Classification of perinatal death in a developing country.

Objectives: To evaluate and compare the three most commonly used perinatal death classification systems: (1) the Nordic–Baltic; (2) the Aberdeen; and (3) the Wigglesworth, and assess their applicability in a developing country (Sudan) with a high perinatal mortality rate, and their justification for practical use in quality assurance and audit activities. Methods: At Omdurman Maternity Hospital (OMH), Khartoum, Sudan, 166 perinatal deaths were prospectively assessed during a 3‐month period (May–August 2000) with a total of 2260 births. Narratives of 166 perinatal deaths were prepared for the purpose of audit. A panel of two Danish and one Sudanese obstetrician categorized the cases according to: (1) the Nordic–Baltic; (2) the Aberdeen; and (3) the Wigglesworth classification. Results: By all three classifications a similar fraction of cases (approx. 85%) were allocated to one category only, and in 15% of cases the assessors were in doubt into which of two categories the cases should be allocated. The necessary information is often not available, giving at least 40% classified as ‘unknown’ in the Aberdeen classification, whereas the Wigglesworth classification results in an even larger group of unspecified asphyxia. Conclusion: Classification of perinatal deaths in developing countries is associated with problems regarding application, validity and usefulness. The Nordic–Baltic classification seems to be most suitable for appropriate stratification using routinely recorded variables and providing categories associated with specific levels of care.

Measles in suburban Khartoum: an epidemiological and clinical study

Clinical and epidemiological data were collected from 187 clinically diagnosed measles patients in Haj Yousif area, suburban Khartoum. Laboratory tests confirmed the diagnosis in 141 (75%) of the cases, but demonstrated that in 46 (25%) patients the clinical symptoms were not caused by an acute measles virus (MV) infection. According to their vaccination card, 59% of the laboratory‐confirmed measles cases had been vaccinated for measles. Compared with non‐measles rash disease cases, confirmed measles cases more often had severe illness (P < 0.0001), were dehydrated (P=0.01) at presentation and less likely to recover without complications [OR 0.19 (95% CI 0.09, 0.39)]. There was no difference in death rate (P=0.20). Underweight [weight‐for‐age Z score (WAZ) ≤ −2 SD] was an independent predictor of recovery with complications [OR 0.4 (95% CI 0.2, 0.99)]. Severe measles cases (those who developed diarrhoea, pneumonia, otitis media, encephalitis or haemorrhagic rash) had similar vaccination rates and time intervals since vaccination as uncomplicated measles cases. Although severe measles had lower WAZ‐scores (P=0.004), none of the nutritional parameters studied were predictive of outcome. Mortality was higher in the severe measles group [OR 8.8 (95% CI 1.7, 85.2)]. In 11 of 141 confirmed measles cases serological evidence of a recent infection with another virus was found, most commonly varicella zoster virus and dengue virus; spotted fever and rubella were among the most frequent diagnoses in 17 of 47 cases of the non‐measles cases.

The role of the village midwife in detection of high risk pregnancies and newborns

The preliminary findings of a prospective study of perinatal, neonatal and maternal mortality carried out in a rural community of Sudan are reported. Out of 6275 deliveries monitored over a period of 3 years, 150 stillbirths, 167 neonatal deaths and 27 maternal deaths were observed. An intervention program to upgrade the skills of the village midwives started in the middle of the second year. There was a 25% reduction in the risk of unfavorable outcome of pregnancy (i.e. stillbirth and neonatal death) in the third year relative to the first 2 years. Peer review of the 40 village midwives who took part in the study revealed their tremendous potentials in mobilization of mothers as well as participation in primary health care. Their role in detection of high risk pregnancies and newborns cannot be overemphasized.

A new complex homozygous large rearrangement of the PINK1 gene in a Sudanese family with early onset Parkinson’s disease

PARK2 and PINK1 gene mutations are involved in recessive early onset Parkinson’s disease (EOPD). In order to determine the causative mutations in three affected sibs from a consanguineous Sudanese family with EOPD, multiplex ligation-dependent probe amplification was performed and revealed that the patients were homozygous for a deletion of PINK1 exons 4 to 8. Breakpoint analysis revealed a complex rearrangement combining a large deletion and the insertion of a sequence duplicated from the DDOST gene intron 2, located near the PINK1 gene. As breakpoint sequences displayed only three base pairs of homology, this rearrangement may result from Fork Stalling and Template Switching mechanism. This third large rearrangement of PINK1 enlarges the mutation spectrum and, together with recent published data in Tunisian patients with EOPD, points out that PINK1 gene analysis, including search for large rearrangement, should be considered in early onset recessive PD patients, particularly those from Arab origin.

Skills retention in Sudanese village midwives 1 year following Helping Babies Breathe training

Background Over 80% of deliveries in Sudan occur in isolated villages, attended by village midwives (VMWs). Upgrading newborn resuscitation skills with the Helping Babies Breathe (HBB) programme could improve newborn survival rates. Objective To describe the competencies in newborn resuscitation of selected VMWs pre-HBB and post-HBB training. Methods In a prospective intervention study, the VMWs’ performances in the HBB Objective Structured Clinical Examination B simulated scenario (manikin requiring face-mask ventilation (FMV)) were digitally recorded and analysed prior to and 3 and 12 months following HBB training. Regular manikin-based practice was encouraged following training. Results Pre-HBB training, 42% of 71 VMWs (of whom 61% were functionally illiterate) stimulated the non-breathing manikin by holding it by the legs and either stimulated/slapped (30.4%) or shook (12.7%) it, while 25% (18/71) provided manikin mouth-to-mouth ventilation. The low scorings on the ‘preparation for birth’ (0% and 3.1% at 3 and 12 months, respectively) were mainly due to failure to demonstrate the subitem of ‘cleans hands’. The percentage of VMWs providing manikin FMV within the Golden Minute increased from 37.3% (25/67) to 72.3% (47/65) (p<0.005), but there were no significant differences in the number of VMWs producing at least five FMVs at 3 months (73%, 49/67) and 12 months (58%, 38/65), respectively. Conclusions VMWs, despite a high illiteracy rate, absorbed and sustained HBB skills for at least a year. Regular, low intensity, manikin-based skills training with peers may have helped sustain FMV, but not hand-cleansing skills.

Prevention of measles in Sudan: a prospective study on vaccination, diagnosis and epidemiology

Despite the availability of safe and effective live attenuated vaccines, measles continues to be endemic in many developing countries. Control and elimination of measles will be especially difficult in East Africa, because of its limited infrastructure and political instability. We have studied diagnostic and epidemiological aspects of measles in suburban Khartoum, Sudan. Prospective studies were carried out in a cohort of clinically diagnosed measles cases and in a cohort of newborns, which were both followed up for 2 years. The studies intended to provide a rational basis for improvement of measles vaccination strategies, and strengthen measles research infrastructure in Khartoum.

Randomized Trials Comparing Inactivated Vaccine after Medium- or High-titer Measles Vaccine with Standard Titer Measles Vaccine after Inactivated Vaccine: A Meta-analysis

Background: Observational studies have suggested that girls have higher mortality if their most recent immunization is an inactivated vaccine rather than a live vaccine. We therefore reanalyzed 5 randomized trials of early measles vaccine (MV) in which it was possible to compare an inactivated vaccines [after medium-titer MV (MTMV) or high-titer MV (HTMV)] and a live standard titer MV (after an initial inactivated vaccine). Methods: The trials were conducted in Sudan, Senegal, The Gambia and Guinea-Bissau. The intervention group received live MTMV or HTMV from 4 to 5 months and then an inactivated vaccine from 9 to 10 months of age; the control children received inactivated vaccine/placebo from 4 to 5 months and standard titer MV from 9 to 10 months of age. We compared mortality from 9 months until end of study at 3 to 5 years of age for children who received inactivated vaccine (after MTMV or HTMV) and standard titer MV (after inactivated vaccine), respectively. The original datasets were analyzed using a Cox proportional hazards model stratified by trial. Results: The mortality rate ratio (MRR) was 1.38 (95% confidence interval: 1.05–1.83) after an inactivated vaccine (after MTMV or HTMV) compared with a standard titer MV (after inactivated vaccine). Girls had a MRR of 1.89 (1.27–2.80), whereas there was no effect for boys, the sex-differential effect being significant (P = 0.02). Excluding measles cases did not alter these conclusions, the MRR after inactivated vaccines (after MTMV or HTMV) being 1.40 (1.06–1.86) higher overall and 1.92 (1.29–2.86) for girls. Control for variations in national immunization schedules for other vaccines did not modify these results. Conclusions: After 9 months of age, all children had been immunized against measles, and mortality in girls was higher when they had received inactivated vaccines (after MTMV or HTMV) rather than live standard titer MV (after an inactivated vaccine).

Perinatal outcomes following Helping Babies Breathe training and regular peer–peer skills practice among village midwives in Sudan

Background Over 80% of deliveries in Sudan occur in rural areas, attended by village midwives (VMWs). Objective To determine the impact of Helping Babies Breathe training and regular peer–peer skills practice (HBBT+RPPSP) on VMW resuscitation practices and outcomes. Methods In a prospective community-based intervention study, 71/82 VMWs, reporting to six East Nile rural medical centres, with previous experience in community health research, consented to HBBT+RPPSP. Outcomes included changes in the resuscitation practices, fresh stillbirths (FSB) and early neonatal deaths <1 week (ENND). Results There were 1350 and 3040 deliveries before and after HBBT+RPPSP, respectively, with no significant differences between the two cohorts regarding maternal age, education or area of birth. Drying of the newborn increased almost tenfold (8.4%, n=113 to 74.9%, n=1011) while suctioning of the mouth/nose decreased fivefold (80.3%, n=2442 to 14.4%, n=437) following HBBT+RPPSP. Pre-HBBT+RPPSP9/18 (50%) newborns who had mouth-to-mouth ventilation died, compared with 13/119 (11%) who received bag-mask ventilation post-HBBT+RPPSP. Excluding 11 macerated fetuses, there were 55 perinatal deaths: 14 FSB/18 ENND (6 months pre-HBBT+RPPSP) and 10 FSB/13 ENND (18 months post-HBBT+RPPSP). FSB rates decreased from 10.5 to 3.3 per 1000 births ((χ2)=8.6209, p=0.003), while ENND rates decreased from 13.5 to 4.3 per 1000 live births ((χ2)=10.9369, p=0.001) pre-HBBT+RPPSP and post-HBBT+RPPSP, respectively. Conclusion In a selected group of VMWs, HBBT+RPPSP was associated with improvements in newborn resuscitation and perinatal outcomes. HBBT+RPPSP could have immense benefits if propagated nationally to all 17 000 VMWs in Sudan.