Volume 29, Number 1—January 2023
Research
Molecular Tools for Early Detection of Invasive Malaria Vector Anopheles stephensi Mosquitoes
Om P. Singh
, Taranjeet Kaur, Gunjan Sharma, Madhavinadha P. Kona, Shobhna Mishra, Neera Kapoor, and Prashant K. Mallick
, Taranjeet Kaur, Gunjan Sharma, Madhavinadha P. Kona, Shobhna Mishra, Neera Kapoor, and Prashant K. Mallick
Table 2
Results of hydrolysis real-time PCR and size-diagnostic PCR on individual and pooled mosquitoes for early detection of invasive malaria vector Anopheles stephensi mosquitoes*
| Specimen type | DNA isolation method | Real-time PCR |
Size-diagnostic PCR |
|||||
|---|---|---|---|---|---|---|---|---|
| No. | Ct values (An. stephensi)† | Ct values IC | No. | An. stephensi– specific band |
IC band | |||
| Anophelines | ||||||||
| Neocellia series | ||||||||
| An. stephensi type | DNeasy | 26 | 13.47–17.23 | 15.20–18.93 | 24 | Positive | Positive | |
| An. stephensi intermediate | DNeasy | 2 | 14.75–15.60 | 15.39–16.06 | 2 | Positive | Positive | |
| An. stephensi var. mysorensis | DNeasy | 1 | 14.54 | 15.01 | 1 | Positive | Positive | |
| An. stephensi strain STE2 | DNeasy | 1 | 14.85–15.25 | 16.06–16.51 | 2 | Positive | Positive | |
| An. stephensi type form | Boiling | 10 | 15.17–17.41 | 15.97–18.05 | - | Not done | Not done | |
| An. stephensi type form, single leg | DNeasy | 4 | 18.64–22.29 | 19.02–23.51 | 4 | Positive | Positive | |
| Pyretophorus series | ||||||||
| An. gambiae | DNeasy | 3 | Negative | 14.58–17.32 | 3 | Negative | Positive | |
| An. quadrimaculatus | DNeasy | 2 | Negative | 14.52–15.64 | 2 | Negative | Positive | |
| An. merus | DNeasy | 2 | Negative | 15.22–15.35 | 2 | Negative | Positive | |
| An. subpictus form A | DNeasy | 2 | Negative | 16.61–17.02 | 1 | Negative | Positive | |
| An. subpictus form A‡ | Pre-isolated§ | 1 | 33.09 | 18.32 | Negative | Positive | ||
| An. subpictus form B | Pre-isolated§ | 2 | Negative | 16.17–16.35 | 1 | Negative | Positive | |
| An. sundaicus cytoform D | Pre-isolated§ | 1 | Negative | 14.47 | 1 | Negative | Positive | |
| Albimanus series | ||||||||
| An. albimanus | DNeasy | 2 | Negative | 16.21–16.58 | 1 | Negative | Positive | |
| Anopheles series | ||||||||
| An. freeborni | DNeasy | 2 | Negative | 15.73–15.81 | 1 | Negative | Positive | |
| An. atroparvus | DNeasy | 2 | Negative | 14.70–15.88 | 1 | Negative | Positive | |
| Neomyzomyia series | ||||||||
| An. dirus | DNeasy | 2 | Negative | 17.34–18.01 | 1 | Negative | Positive | |
| An. farauti | DNeasy | 2 | Negative | 19.09–19.48 | 1 | Negative | Positive | |
| Myzomyia series | ||||||||
| An. funestus | DNeasy | 2 | Negative | 15.50–15.62 | 1 | Negative | Positive | |
| An. culicifacies species A | DNeasy | 1 | Negative | 17.41 | 1 | Negative | Positive | |
| An. culicifacies species B | Pre-isolated§ | 1 | Negative | 17.48 | 1 | Negative | Positive | |
| An. fluviatilis species S | Pre-isolated§ | 1 | Negative | 16.61 | 1 | Negative | Positive | |
| An. fluviatilis species T |
Pre-isolated§ |
1 |
Negative |
17.02 |
1 |
Negative |
Positive |
|
| Culicines | ||||||||
| Culex quinquefasciatus | DNeasy | 3 | Negative | 16.58–19.20 | 2 | Negative | Positive | |
| Aedes albopictus | DNeasy | 2 | Negative | 16.65–16.68 | 2 | Negative | Positive | |
| Ae. aegypti | DNeasy | 2 | Negative | 18.96–19.20 | 2 | Negative | Positive | |
| Ae. aegypti‡ |
Pre-isolated§ |
1 |
32.95 |
20.97 |
Negative |
Positive |
||
| Single An. stephensi individual pooled with An. culicifacies¶ | ||||||||
| Adults (1/2) | DNeasy | 1 | 17.06 | 18.31 | 1 | Positive | Positive | |
| Adults (1/5) | DNeasy | 1 | 15.37 | 16.11 | 1 | Positive | Positive | |
| Adults (1/10) | DNeasy | 1 | 15.77 | 15.30 | 1 | Positive | Positive | |
| Adults (1/20) | DNeasy | 1 | 15.74 | 13.71 | 1 | Positive | Positive | |
| Adults (1/50) | DNeasy | 1 | 15.43 | 11.43 | 1 | Positive | Positive | |
| Adults (1/25) | DNAzol | 1 | 14.53 | 12.61 | 1 | Positive | Positive | |
| Adults (1/100) | DNAzol | 1 | 16.37 | 11.27 | 1 | Positive | Positive | |
| Adults (1/500) | DNAzol | 2 | 20.12–23.96 | 11.08–11.50 | 2 | Positive | Positive | |
| Larvae (1/2) | DNeasy | 1 | 15.56 | 16.87 | 1 | Positive | Positive | |
| Larvae (1/5) | DNeasy | 1 | 15.49 | 16.65 | 1 | Positive | Positive | |
| Larvae (1/10) | DNeasy | 1 | 16.36 | 15.53 | 1 | Positive | Positive | |
| Larvae (1/20) | DNeasy | 1 | 16.51 | 15.06 | 1 | Positive | Positive | |
| Larvae (1/50) | DNeasy | 1 | 16.27 | 14.40 | 1 | Positive | Positive | |
| Larvae (1/100) |
DNAzol |
1 |
18.14 |
13.16 |
1 |
Positive |
Positive |
|
| Single An. stephensi individual pooled with field collected mosquitoes (mixed mosquito-species)¶ | ||||||||
| Adults (1/100) | DNAzol | 1 | 17.61 | 11.70 | 1 | Positive | Positive | |
*Ct, cycle threshold; IC, internal control. †Ct value shown as Negative implies that no Ct value was scored within 35 cycles of reactions. ‡Contaminated with An. stephensi DNA. §Preserved DNA isolated manually for other studies. ¶Denominator of numeric expression inside parenthesis indicate total number of mosquitoes in a pool.
Page created: November 10, 2022
Page updated: December 21, 2022
Page reviewed: December 21, 2022
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