Born in Bila Tserkva on July 12, 1916, Pavlichenko moved to Kiev with her family at the age of fourteen. There she joined a shooting club and developed into a sharpshooter, while working as a grinder at the Kiev Arsenal factory

There she became one of 2,000 female snipers in the Red Army, of whom only about 500 ultimately survived the war. As a sniper, she made her first two kills near Belyayevka, using a Mosin-Nagant bolt action rifle with a P.E. 4-power scope.

Pvt. Pavlichenko fought for about two and a half months near Odessa, where she recorded 187 kills. When the Germans gained control of Odessa, her unit was pulled to be sent to Sevastopol on the Crimean Peninsula. In May 1942, Lieutenant Pavlichenko was cited by the Southern Army Council for killing 257 German soldiers. Her total confirmed kills during World War II was 309, including 36 enemy snipers. In June 1942, Pavlichenko was wounded by mortar fire. Because of her growing status, she was pulled from combat less than a month after recovering from her wound.

She was sent to Canada and the United States for a publicity visit and became the first Soviet citizen to be received by a U.S. President when Franklin Roosevelt welcomed her at the White House. Later, Pavlichenko was invited by Eleanor Roosevelt to tour America relating her experiences. She appeared before the International Student Assembly being held in Washington, D.C., and later attended CIO meetings and made appearances and speeches in New York City. The United States gave her a Colt automatic pistol, and in Canada, she was presented with a sighted Winchester rifle, the latter of which is now on display at the Central Museum of the Armed Forces in Moscow. While visiting in Canada along with Vladimir Pehelintsev (fellow sniper) and Nikolai Krasavchenko (Moscow fuel commissioner), they were greeted by thousands at Toronto's Union Station.

Having attained the rank of Major, Pavlichenko never returned to combat but became an instructor and trained Soviet snipers until the war's end. In 1943, she was awarded the Gold Star of the Hero of the Soviet Union, and was commemorated on a Soviet postage stamp.