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. 2014 Sep-Oct;21(5):17-23.

Comparison of nested polymerase chain reaction and microscopy as diagnostic tools in congenital malaria: a study at tjark corneile hillers hospital maumere, indonesia

Natalia Erica Jahja  1 , Irene Ratridewi Huwae  1 , Mario B Nara  2 , Adinda Harley  3 , Tarina Widaningrum  4 , Loeki Enggar Fitri  5
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Comparison of nested polymerase chain reaction and microscopy as diagnostic tools in congenital malaria: a study at tjark corneile hillers hospital maumere, indonesia

Natalia Erica Jahja et al. Malays J Med Sci. 2014 Sep-Oct.

Abstract

Background: Microscopic detection is the conventional method for detecting malaria parasites. Although it is efficient and inexpensive, it has its limitations. In recent years, polymeras chain reaction (PCR) has been considered superior to microscopy in detecting mixed Plasmodium infections or infections with low parasite density. To determine whether microscopic or nested PCR (nPCR) is better at detecting congenital malaria (CM).

Methods: Blood smear examination and nPCR were performed with blood samples taken from mothers and their newborns, who were likely to be suffering from CM and in whom one of the symptoms was low birth weight (LBW). The sensitivity and specificity of each method were then compared.

Results: During one year of study, the prevalence of CM among 92 LBW newborns was determined to be 6.8% using microscopy and 7.8% using nPCR. Among the 92 mother-infant paired subjects, CM was detected in 34 subjects (37%) by microscopy and in 39 subjects (42.4%) by nPCR. nPCR was more sensitive (76.5% vs 66.7%) but less specific (77.6% vs 84.9%) than microscopy. When the two methods were compared, nPCR gave significantly better results in diagnosing CM (AUC = 0.770; P < 0.001).

Conclusion: Although microscopy remains the most appropriate method for the diagnosis of CM in remote areas, nPCR can be considered a complementary test.

Keywords: congenital; malaria; microscopy; nested PCR; newborns.

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Figures

Figure 1:
Figure 1:
Example of comparative identification of Plasmodium species using microscopy and nPCR. a) Identification by microscopy showed trophozoites/ring form on enlarged red blood cell (red arrow). Active macrophage ingesting Plasmodium is showed by blue arrow; b) Confirmation by PCR showed mixed infection of P. falciparum (upper gel, 1st column, 205 bp) and P. vivax (lower gel, 1st column, 117 bp). c) Identification by microscopy showed trophozoites/ring form on enlarged red blood cell (white arrow) but there was an image resembling schizont of P. falciparum on upper field (brown arrow); d) Confirmation by PCR showed single infection of P. vivax (lower gel, 12th column, 117 bp).
Figure 2:
Figure 2:
Summary of Receiver Operating Characteristic (ROC) plot of sensitivity and specificity of microscopy and nPCR. a) ROC plot of microscopy method (blue line) using PCR (green line) as a gold standard and AUC obtained was 0.758 (P < 0.001). b) ROC plot of nPCR method (blue line) using microscopy (green line) as a gold standard and AUC obtained was 0.770 (P < 0.001).
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